Moisturizing antioxidant composition and application thereof in cosmetics

Abstract

The invention provides a moisturizing and antioxidant composition and application thereof in cosmetics. According to the invention, the HA-hexapeptide-9 derivative, the fritillaria aureocauda bulb extract and the semen cuscutae extract are compounded to prepare the moisturizing and antioxidant composition, and the composition can achieve the effects of resisting oxidation and moisturizing by improving the expression of GSH, SOD, UCA and PCA. The composition is used as a main effective component to prepare skin care cosmetics. Through human body efficacy experiment tests, the skin care product has the effects of improving skin moisture content, skin elasticity and corium layer thickness and density.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of cosmetics, and particularly relates to a moisturizing antioxidant composition and application thereof in cosmetics. BACKGROUND

[0002] The information disclosed in the background of the present application is merely intended to increase the understanding of the overall background of the present application and should not be necessarily regarded as acknowledging or implicitly suggesting that this information constitutes prior art known to those of ordinary skill in the art.

[0003] Free radicals, also known as reactive oxygen species (ROS), are compounds formed when an oxygen molecule combines with another molecule to produce an odd number of electrons. Oxygen molecules with paired electrons are stable; however, oxygen with unpaired electrons is "reactive" because it will seek out and steal electrons from important components, damaging them. DNA, cytoskeletal elements, cellular proteins, and cell membranes can all be adversely affected by reactive oxygen species. ROS are not only involved in the overall aging process, but also cause skin photoaging, carcinogenesis, and inflammation. Free radicals also play an important role in intrinsic and extrinsic skin aging.

[0004] The body has developed a defense mechanism known as antioxidants, which prevent the damage caused by free radicals by reducing and neutralizing them. Naturally occurring antioxidant enzymes in the skin include superoxide dismutase (SOD), catalase, and glutathione peroxidase (GPx); non-enzymatic endogenous antioxidant molecules are alpha-tocopherol (vitamin E), ascorbic acid (vitamin C), glutathione (GSH), and ubiquinone. As part of the natural aging process, our body's defense mechanisms weaken while the production of ROS increases. This leads to an imbalance and an uncontrolled increase in the number of free radicals that damage DNA, cytoskeletal elements, cellular proteins, and cell membranes.

[0005] Filaggrin (FLG) is a key protein for the skin to perform its moisturizing function, and FLG monomers are degraded into free amino acids, including glutamine, arginine, and histidine, and then converted into urocanic acid (UCA) and pyrrolidone carboxylic acid (PCA). This process is mediated by other groups of proteases, including Caspase 14, calpain 1, and bleomycin hydrolase. PCA is the main component of natural moisturizing factors (NMFs), which is responsible for maintaining moisture in the stratum corneum, thereby enhancing the skin's moisturizing function.

[0006] Antioxidants are a class of substances that can help capture and neutralize free radicals, thereby eliminating the damage of free radicals to the human body. The currently widely used antioxidant ingredients have certain shortcomings, which may be related to their own properties, and are easily oxidized. Most of them are unstable in aqueous solution. For example, the external antioxidant VC component has greater irritation, and at the same time, the stability is poor. Directly using glutathione and SOD enzyme in the formula has poor stability, or is easy to have odor, and has high requirements for the formulation or form of the formula. The VE component is an oil-soluble component, and the penetration is stronger than that of VC, but the skin feel is sticky, and the skin feel is poor when the addition amount is large. There is an urgent need for stable, efficient and highly compatible antioxidant ingredients. SUMMARY

[0007] In view of the shortcomings of the prior art, the present application provides a moisturizing antioxidant composition and its application in cosmetics. The moisturizing antioxidant composition is prepared by compounding HA-hexapeptide-9 derivative, fritillaria imperialis bulb extract and cuscuta extract. The above-mentioned composition can improve the expression of GSH, SOD, UCA and PCA, thereby achieving the effects of antioxidant and moisturizing. Then, the composition is used as the main effective component to prepare a skin care cosmetic. Through human efficacy experiment test, it has the effects of improving skin water content, skin elasticity, dermal layer thickness and density.

[0008] In order to achieve the above technical purpose, the technical scheme provided by the present application is as follows: a moisturizing antioxidant composition, the moisturizing antioxidant composition comprises the following components in mass fraction: HA-hexapeptide-9 derivative 0.05-1.0 parts, fritillaria imperialis bulb extract 1.0-5.0 parts, and cuscuta extract 1.0-10.0 parts.

[0009] Further, the moisturizing antioxidant composition comprises the following components in mass fraction: HA-hexapeptide-9 derivative 0.1-0.7 parts, fritillaria imperialis bulb extract 1.5-4.0 parts, and cuscuta extract 5.0-8.0 parts.

[0010] The preparation method is as follows: mixing HA-hexapeptide-9, fritillaria imperialis bulb extract and cuscuta extract, and dissolving them in purified water (supplemented to 100 parts).

[0011] The structure formula of the HA-hexapeptide-9 derivative is as follows, which can be obtained by market purchase or by the following method (patent application number: 2025111030013).

[0012] Formula (I): ; Wherein .

[0013] The Fritillaria cirrhosa bulb extract is obtained by a commercially available method or the following method, and the preparation method comprises the following steps: adding the Fritillaria cirrhosa bulb dry powder into 65%-80% ethanol solution, heating in a water bath at 60-80 DEG C for 2-4 times, combining the filtrates, concentrating under reduced pressure, volatilizing until no alcohol smell, adding 0.7-1.2 times mass of butanediol, stirring uniformly, and filtering to obtain the Fritillaria cirrhosa bulb extract. The concentration is 0.25-0.40 g / mL of crude drug, that is, 0.25-0.40 g of dry Fritillaria cirrhosa bulb powder is used to obtain 1 mL of Fritillaria cirrhosa bulb extract (solution).

[0014] The Cuscuta extract is obtained by a commercially available method or the following method, and the preparation method comprises the following steps: taking the Cuscuta powder to obtain an extraction liquid by using ultrasonic-assisted subcritical extraction technology, filtering, and adding glycerol after concentration to obtain the Cuscuta extract.

[0015] Specifically, the preparation method comprises the following steps: S1: the Cuscuta powder is added into water for ultrasonic treatment, wherein the material-liquid mass ratio is 1:5-1:10, the power of the ultrasonic wave is set to 300-500 W, the temperature is 25-40 DEG C, and the time is 25-40 min; S2: the material liquid prepared in step S1 is placed in an extractor, 0.5-1.0 times mass of water of the material liquid prepared in S1 is added, the extraction conditions are as follows: the temperature is 120-140 DEG C, the pressure is 0.1-0.8 MPa, the time is 30-60 min, the extraction liquid is filtered, the filtrate is concentrated under reduced pressure, 1-1.5 times mass of glycerol of the concentrated liquid is added, the mixture is stirred and mixed uniformly, and the Cuscuta extract is obtained after filtration, and the concentration of the extraction liquid is 0.30-0.45 g / mL (calculated according to the amount of crude drug).

[0016] The above composition can improve the expression of GSH, SOD, UCA and PCA, thereby achieving the effects of antioxidation and moisturization.

[0017] Therefore, in a second aspect of the present application, the above moisturizing and antioxidant composition is provided for use in the preparation of cosmetics. Obviously, the cosmetics have the effects of antioxidation, anti-aging, improvement of facial wrinkles and skin moisture content.

[0018] In a third aspect of the present application, a cosmetic having the effects of antioxidation, anti-aging, improvement of facial wrinkles and skin moisture content is provided, and the cosmetic at least comprises the above moisturizing and antioxidant composition.

[0019] According to the cosmetic of the present application, the addition amount of the moisturizing and antioxidant composition is 1-35% (w / w), such as 1%, 3%, 5%, 7%, 10%, 15%, 17%, 20%, 25%, 30%, 35%, etc., of the total mass of the cosmetic, which is not limited herein.

[0020] The cosmetic product can also comprise other raw material components permitted to be added in the cosmetic field, including but not limited to emulsifiers, emollients, humectants, cooling agents, skin conditioning agents, thickening agents, preservatives, and the like.

[0021] Meanwhile, the present application can also prepare different cosmetic dosage forms such as essence water, essence milk, essence lotion, shampoo, hair conditioner, hair mask, and the like by reasonably adding the above raw material components, which are not specifically limited herein.

[0022] Technical effects of the present application: 1. The composition prepared by compounding HA-hexapeptide-9 derivative, fritillaria cirrhosa bulbus extract, and semen cuscutae extract can significantly promote the expression of SOD, GSH, PCA, and UCA, thereby achieving the effects of effective moisturizing and antioxidant, and greatly improving the skin care efficacy.

[0023] 2. When HA-hexapeptide-9 derivative, fritillaria cirrhosa bulbus extract, and semen cuscutae extract are combined, the discoloration problem of fritillaria cirrhosa bulbus extract under high temperature and light can be significantly improved, and the stability of the formula can be greatly improved.

[0024] 3. The skin care cosmetic prepared by using the moisturizing and antioxidant composition of the present application has a simple and easy-to-operate preparation method and saves cost. Through human efficacy experiment test, the cosmetic containing the moisturizing and antioxidant composition has the effects of improving skin water content, skin elasticity, dermal layer thickness, and density, and therefore has good moisturizing, anti-aging, and skin elasticity enhancing effects. BRIEF DESCRIPTION OF DRAWINGS

[0025] Figure 1 Figure 1 is a picture showing the improvement of skin dermal density state before and after using the sample for No. 1, No. 28, and No. 32 subjects. DETAILED DESCRIPTION

[0026] It should be noted that the following detailed description is exemplary and is intended to provide further explanation of the present application. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the present application belongs.

[0027] The present application will be further described in conjunction with specific examples, which are only intended to explain the present application and do not limit the content thereof. If the specific conditions of the experiments are not specified in the examples, the general conditions or the conditions recommended by the reagent companies are usually used; the reagents, consumables, and the like used in the following examples can be obtained from commercial channels if not specifically stated.

[0028] The specific preparation method of the HA-hexapeptide-9 derivative in the embodiments of the present application is as follows: S1: Dissolve sodium hyaluronate (trade name: Hua Ling 400, Shandong Jiaofeng Furuida Biological Co., Ltd.) in 0.1 M PBS buffer (pH 7.0) to prepare a solution with a concentration of 1 mM; dissolve hexapeptide-9 (purity ≥98%) in 0.1 M PBS buffer (pH 7.0) to prepare a solution with a concentration of 5 mM.

[0029] S2: Add the alkyne amide condensing agent to the HA solution, wherein the molar ratio of the alkyne amide condensing agent to the carboxyl group of HA is 1:1.2, and stir at room temperature (25°C) for 30 minutes to activate the carboxyl group (-COOH) of HA to form a higher-activity enol ester intermediate (preferably the alkyne amide condensing agent is R1 = Me, EWG = Ts).

[0030] S3: Slowly add the hexapeptide-9 solution prepared in S1 to the activated HA solution (dropping speed 5 mL / min), and continue to stir for 4 hours.

[0031] S4: After the reaction is completed, remove the unreacted alkyne amide condensing agent, free hexapeptide-9, and condensation by-product acetamide derivative by centrifugation (3000 rpm, 4°C) through an ultrafiltration tube with a molecular weight cutoff of 1000 Da; collect the cutoff liquid, and freeze-dry (-50°C, vacuum degree 10 Pa) to obtain the HA-hexapeptide-9 derivative.

[0032]

[0033] The fritillaria cirrhosa bulb extract in the examples and comparative examples of the present application is prepared by the following method: an appropriate amount of fritillaria cirrhosa bulb dry powder is taken, 70% ethanol aqueous solution is added at a solid-liquid ratio of 1:10, heated in a water bath at 70-80°C for 3h, repeated twice, the filtrates are combined, the filtrate is concentrated under reduced pressure, and volatilized to remove alcohol, butanediol is added in an amount equal to that of the concentrated solution, stirred uniformly, and filtered to obtain the fritillaria cirrhosa bulb extract. The concentration is 0.30g / mL in crude drug, that is, 0.30g of dry fritillaria cirrhosa bulb powder is used to obtain 1mL of fritillaria cirrhosa bulb extract (solution).

[0034] In the examples and comparative examples of the present application, the cuscuta extract is prepared by the following method: The specific steps of the preparation method of the cuscuta extract are as follows: S1: The dried cuscuta is crushed through a 50-mesh sieve, an appropriate amount of cuscuta powder is accurately weighed, added to water, the solid-liquid mass ratio is 1:7, the power of the ultrasonic wave is set to 400W, the temperature is 25°C, and the time is 30 min, and after ultrasonic completion, it is used.

[0035] S2: The solution prepared in S1 was placed in an extractor, and 0.8 times the mass of water of the solution prepared in S1 was added. The extraction conditions were a temperature of 130°C, a pressure of 0.5 MPa, and a time of 40 min. The extraction solution was filtered, and the filtrate was concentrated under reduced pressure to about 1 / 15 of the original volume. Glycerol was added to the concentrated solution in an amount of 1.5 times the volume of the concentrated solution, and the mixture was stirred until uniform. After filtration, the extract of Semen Cuscutae was obtained, and the concentration of the extract was about 0.40 g / mL (calculated based on the amount of crude drug).

[0036] Examples 1-3 and Comparative Examples 1-9: The specific schemes are shown in Table 1, and the amounts are calculated by mass. The preparation method was as follows: HA-hexapeptide-9, Fritillaria verticillata bulb extract, and Semen Cuscutae extract were mixed according to the amounts used in each example and comparative example, and were dissolved in purified water.

[0037] Table 1: List of ingredients for specific examples and comparative examples

[0038] Example 4: Moisturizing and antioxidant face cream The above composition was used to prepare a face cream product containing the moisturizing and antioxidant composition, and the formula is shown in Table 2.

[0039] Table 2: Formula of the moisturizing and antioxidant face cream product

[0040] Preparation method of the moisturizing and antioxidant face cream: S1: The raw materials were added to Phase A, respectively, and heated to 80-85°C for stirring and dissolution, and were kept for later use. The raw materials were added to Phase B, respectively, and heated to 75-80°C for stirring and melting, and were kept for later use; S2: The raw materials of Phase B were added to Phase A, and stirring and homogenization were started, and were homogenized for 3-10 min, and then the temperature was lowered; S2: When the temperature was lowered to about 65-70°C, the ingredients of Phase C were added, and stirring was started for dispersion, dissolution, and homogenization, and after dissolution was completed, the temperature was continued to be lowered; S3: When the temperature was lowered to 40-45°C, the ingredients of Phase D were added, and stirring was started for dissolution and dispersion, and the face cream product was obtained.

[0041] Test Example 1: Irritation test experiment The product was subjected to eye irritation testing, i.e., safety testing, according to SN / T 2329-2009 "Cosmetic Eye Irritation / Chicken Embryo Chorioallantoic Membrane Test".

[0042] Experimental method: Fertilized chicken embryos less than 7 days old were incubated in an incubator at 37.6±0.5°C and 50%-70% humidity for 9 days.

[0043] CAM preparation: carry out egg inspection, mark the position of the gas chamber on the eggshell surface, use tweezers to peel off the marked eggshell part, expose the white egg membrane, and operate carefully without damaging the integrity of the egg membrane. Drop 0.5 mL of 0.9% NaCl solution to fully wet the egg membrane, gently absorb the surface liquid with a paper towel, and carefully remove the inner membrane with tweezers to ensure that the blood vessel membrane is not damaged.

[0044] Reaction time method: take 0.3 mL of transparent test substance and evenly drop or apply it on the surface of the CAM, and after 3 min of action, wash away the test substance with 0.9% NaCl solution, observe the degree of change of each toxicity effect of the CAM, and record the time of occurrence of each toxicity effect within 5 min of action. 6 chicken embryos are set for each test substance, and 1 chicken embryo is set for negative control (normal saline) and positive control (1% SDS). The scoring criteria and phenomena are shown in Table 3.

[0045] Table 3 Scoring criteria and phenomena

[0046] Stimulation scoring method: the reaction time method is used for the test, and the stimulation score (IS) is calculated using the following formula, and the results are rounded to two decimal places:

[0047] In the formula: sec H (blood vessel melting time) - the average time observed on the CAM membrane for the onset of blood vessel melting, in seconds (s); sec L (coagulation time) - the average time observed on the CAM membrane for the onset of coagulation, in seconds (s). sec C (coagulation time) - the average time observed on the CAM membrane for the onset of coagulation, in seconds (s).

[0048] According to the IS value, the eye irritation of the test substance is classified according to Table 4.

[0049] Table 4 Evaluation of stimulation scoring method results

[0050] The test results are shown in Table 5, and the moisturizing antioxidant composition has no irritation.

[0051] Table 5 Statistical table of irritation test results

[0052] Test Example 2: GSH and SOD expression experiment The experiment was carried out with EpiKutis® as a model cell strain. According to the test grouping (BC, NC and PC groups) in Table 3, the model was transferred to a 6-well plate (0.9 mL of EpiGrowth (Guangdong Boxi Shanxi Branch) culture solution was added in advance), and the test group number was marked on the 6-well plate. The BC and NC groups were not treated, and the PC group was added with 0.9 mL of working solution of corresponding concentration, and the sample group was evenly distributed on the surface of the model.

[0053] UVB irradiation: all groups were subjected to UVB irradiation except the BC group, the irradiation dose was 600 mJ / cm 2 , the irradiation time was about 25 min, and after the irradiation was completed, the 6-well plate was placed in a CO2 incubator (37℃, 5% CO2) for 24 h.

[0054] Immunofluorescence detection: the model for detection was fixed with 4% paraformaldehyde, and after 24 h of fixation, immunofluorescence detection was performed, and the pictures were collected and analyzed under a fluorescence microscope.

[0055] ELISA test: after incubation, the 3D epidermal skin model culture solution was collected in a centrifuge tube, and after collection, the sample for ELISA detection was stored in a-80℃ refrigerator, and detection and analysis were performed according to the operation instruction of the ELISA kit.

[0056] Chemical colorimetric method test: the model for detection was quickly frozen in liquid nitrogen, and after being ground into powder, detection was performed according to the operation instruction of the GSH kit.

[0057]

[0058] GraphPad Prism was used for plotting, and the results were expressed as Mean±SD. The comparison between groups was analyzed by t -test. The statistical analysis was two-tailed. 0.01<0.05 was considered to have significant difference, P <0.01 was considered to have extremely significant difference. Compared with the NC group, significance*indicated that P value<0.05 was indicated as*, P- value<0.01 was indicated as**. P-

[0059] The test scheme is shown in Table 6, and the test results are shown in Tables 7-8.

[0060] Table 6 Test scheme

[0061] ​Note: sample concentration 5 mg / mL (diluted with deionized water).

[0062] Table 7 SOD content detection result summary table

[0063] Note: ## indicates compared with blank control, p <0.01; ** indicates compared with blank control, p <0.01; * indicates compared with blank control, 0.01 p <0.05.

[0064] As can be seen from Table 7, compared with the NC group, the SOD content of the PC group increased significantly, indicating that the positive control test was effective. Single fritillariae cirrhosae bulb extract (Comparative Example 2) did not have the effect of promoting SOD synthesis, and Examples 1-3 could significantly increase the SOD content, indicating that the HA-hexapeptide-9 derivative could have a significant synergistic promoting effect with fritillariae cirrhosae bulb extract and semen cuscutae extract. Comparative Example 3, single semen cuscutae extract, did not have a significant effect on promoting SOD content. As can be seen from Comparative Example 4, the HA-hexapeptide-9 derivative did not have a significant synergistic effect with fritillariae cirrhosae bulb extract. Comparative Example 9, a simple physical mixture of HA and hexapeptide-9 mixture, did not have as obvious a synergistic effect as Examples 2 and 3 with fritillariae cirrhosae bulb extract and semen cuscutae extract.

[0065] Table 8 GSH content detection result summary table

[0066] Note: ## indicates compared with blank control, p <0.01; ** indicates compared with blank control, p <0.01; * indicates compared with blank control, 0.01 p <0.05.

[0067] As can be seen from Table 8: compared with the NC group, the GSH content of the PC group increased significantly, indicating that the positive control test was effective this time. Examples 1-3 can significantly improve the GSH content, indicating that the HA-hexapeptide-9 derivative can have a significant synergistic promotion effect with Fritillariae Liliaceae Bulbus extract and Cuscuta Japonica Extract. Single Cuscuta Japonica Extract does not have the effect of promoting the generation of GSH. Single Fritillariae Liliaceae Bulbus extract has a certain effect of promoting the generation of GSH, but the effect is not significant. As can be seen from Comparative Example 4 and Comparative Example 5, the HA-hexapeptide-9 derivative has a certain synergistic effect with Fritillariae Liliaceae Bulbus extract and Cuscuta Japonica Extract, but the synergistic effect is not as obvious as that of Examples 2 and 3. The synergistic effect of the simple physical mixture of HA and hexapeptide-9 mixture with Fritillariae Liliaceae Bulbus extract and Cuscuta Japonica Extract in Comparative Example 9 is not as obvious as that of Examples 1-3.

[0068] Test Example 3: PCA and UCA content detection experiment The experiment was carried out with EpiKutis® as a model for experimental research. According to the test scheme in Table 9, EpiKutis® was transferred to a 6-well plate (0.9 mL of EpiGrowth culture solution (Guangdong Boxi Shanxi Branch) was added in advance). The NC group, the PC group and the sample group were added with 25 μL of 0.1% SLS solution on the surface, and incubated for 30 min. After incubation, the PC group was added with the corresponding working solution, and the sample group was evenly distributed with the sample working solution (12.5 μL) on the surface of the model, and incubated in a CO2 incubator (37°C, 5% CO2) for 24 h. After incubation, the residual test substance was washed with sterile PBS solution, and the residual liquid was wiped off with a sterile cotton swab. The EpiKutis® model was cut into a 1.5 mL centrifuge tube, 500 μL of 0.2 mg / mL proteinase K was added to each tube, and it was placed in a 50°C water bath for 1 h. After the cuticle was removed, 250 μL of methanol was added to each tube, and it was ultrasonicated for 30 min, and then centrifuged at 14000 rpm for 10 min. After the methanol was evaporated at 60°C, high performance liquid chromatography analysis was performed, and the results are shown in Tables 10-11.

[0069]

[0070] GraphPad Prism was used for plotting, and the results were expressed as Mean±SD. The comparison between groups was analyzed by t -test statistical analysis. The statistical analysis was two-tailed. 0.01< P< 0.05 was considered to have a significant difference, P <0.05 was considered to have a significant difference, P <0.01 was considered to have a very significant difference. Compared with the NC group, significant*indicates, P- value<0.05 was considered to have a significant difference, P- value<0.01 was considered to have a very significant difference.

[0071] Table 9 Test scheme

[0072] Note: Sample concentration 5 mg / mL (diluted with deionized water).

[0073] Table 10 PCA detection result summary table

[0074] Note: ## indicates compared with blank control, p <0.01; ** indicates compared with blank control, p <0.01; * indicates compared with blank control, 0.01 p <0.05.

[0075] As can be seen from Table 10: compared with the NC group, the PCA content of the PC group increased significantly, indicating that the positive control test was effective this time. Examples 1-3 can significantly improve the PCA content, indicating that the HA-hexapeptide-9 derivative can have a significant synergistic promotion effect with Fritillariae cirrhosae Bulbus extract and Cuscuta japonica extract. Single Cuscuta japonica extract does not have the effect of promoting PCA generation. As can be seen from Comparative Example 4 and Comparative Example 5, the HA-hexapeptide-9 derivative has a certain synergistic effect with Fritillariae cirrhosae Bulbus extract and Cuscuta japonica extract, but not as significant as Examples 2-3. As can be seen from Comparative Example 6, Fritillariae cirrhosae Bulbus extract and Cuscuta japonica extract have a certain synergistic promotion effect, and Comparative Example 9 shows that the synergistic effect of the simple physical mixture of HA and hexapeptide-9 mixture with Fritillariae cirrhosae Bulbus extract and Cuscuta japonica extract is not as obvious as the synergistic effect of Examples 2 and 3.

[0076] Table 11 UCA content detection result summary table

[0077] Note: ## indicates compared with blank control, p <0.01; ** indicates compared with blank control, p <0.01; * indicates compared with blank control, 0.01 p <0.05.

[0078] As can be seen from Table 11, the UCA content of the PC group increased significantly compared with the NC group, indicating that the positive control test was effective. Examples 1-3 can significantly increase the UCA content, indicating that the HA-hexapeptide-9 derivative can have a significant synergistic promoting effect with the Fritillariae Thunbergii Miq Bulbus extract and the Cuscuta Chinensis Lam extract. Single Fritillariae Thunbergii Miq Bulbus extract does not have the effect of promoting the generation of UCA. As can be seen from Comparative Example 4 and Comparative Example 5, the HA-hexapeptide-9 derivative also has a certain synergistic effect with the Fritillariae Thunbergii Miq Bulbus extract and the Cuscuta Chinensis Lam extract. The synergistic effect of the simple physical mixture of HA and hexapeptide-9 mixture with the Fritillariae Thunbergii Miq Bulbus extract and the Cuscuta Chinensis Lam extract is not as obvious as that of Examples 2 and 3.

[0079] Test Example 4: Stability test experiment The examples and comparative examples were subjected to stability tests and were placed in cold-heat cycles (-18℃, 4℃, 45℃, cycled once every 3 days) for 4 weeks, 45℃, -18℃ and 4℃ for 12 weeks. Observation was carried out every two weeks, and the samples were cooled to room temperature at the time of observation. The stability of the product was judged by whether color change, taste change, precipitation and other phenomena occurred. If no color change, taste change, precipitation and other unstable phenomena occurred, the product was stable. The color deepening situation was described as follows: the larger the number, the more serious the discoloration. First level: o, second level: o o, third level: o o o. The precipitation situation was described as follows: the larger the number, the more precipitates, first level: Δ, second level: ΔΔ, third level: ΔΔΔ.

[0080] Table 12: Stability test results

[0081] As can be seen from Table 12, the Fritillariae Thunbergii Miq Bulbus extract alone is prone to discoloration under light, 45℃, cold-heat cycle conditions, and is prone to precipitation at -18℃. The Fritillariae Thunbergii Miq Bulbus extract mixed with the HA-hexapeptide-9 physical mixture and the Cuscuta Chinensis Lam extract did not completely improve the discoloration and precipitation problems, but when the HA-hexapeptide-9 derivative was mixed with the Fritillariae Thunbergii Miq Bulbus extract and the Cuscuta Chinensis Lam extract, the discoloration and precipitation problems were well improved, greatly improving the stability of the formula.

[0082] Test Example 5: Skin feel test experiment The examples 1~3 and VE were respectively subjected to skin feel test. The scores were given according to the smoothness during the application process and the stickiness after use. The smoothness (the index of the degree of skin stickiness and slip) score standard: 1-10, the higher the score, the smoother, the lower the score, the stickier. The stickiness (after use, the test product remains on the skin surface and causes the skin to feel sticky and resistant) score standard: 1-10, the higher the score, the drier, the lower the score, the stickier. 15 testers were selected, and the examples 1~3 and VE were respectively subjected to application test, and finally the average score was obtained. The test results are shown in Table 13.

[0083] Table 13 Skin feel test results

[0084] As can be seen from Table 13, in terms of smoothness, the examples 1~3 are smoother, and the smoothness of VE is far lower than that of the examples 1~3. In terms of stickiness, the examples 1~3 are drier, and the dryness of VE is far lower than that of the examples 1~3. It also shows that VE is stickier than the examples 1~3.

[0085] Test example 5: evaluation of human body moisturizing and anti-aging efficacy The sample size of 34 people was selected, including 31 women and 3 men, and the age of the subjects was 27-45 years old, and the test area was the face. The detection time: before using the sample (D0), after using the sample for 14 days (D14), and after using the sample for 28 days (D28). Test instrument: Cutometer (MPA580, Courage and Khazaka, Germany) was used to detect skin elasticity and tightness, DUB Skin Scanner (DUBcutis, TPM, Germany) was used to quantify the dermal thickness and dermal density, and was used to evaluate the improvement of the skin tightness. The skin moisture content tester Corneometer (CM 825, Courage and Khazaka, Germany) was based on the principle of capacitance to test the skin moisture content, and was used to detect the relative water content on the skin surface, and the higher the value, the higher the water content.

[0086] Experimental parameters: Skin elasticity: the larger the R2, R5 and R7 values of skin elasticity, the better the skin elasticity.

[0087] Dermal thickness test: the larger the dermal thickness value, the tighter the skin.

[0088] Initial value: refers to the basic value when no sample is used.

[0089] Average value = , wherein x is the individual parameter measurement value, and n is the number of effective data.

[0090] Difference from initial value T使用后 - T使用前 .

[0091]

[0092] Statistical analysis was performed using SPSS Statistics 25, two-tailed test, test level a = 0.05.

[0093] According to the normal distribution test results, the difference analysis method is selected: if the measured value is normally distributed, the t test method is used for statistical analysis; if it is not normally distributed, the rank sum test method is used for statistical analysis.

[0094] The test results are shown in Tables 14-19. The improvement of the skin dermal density state of subjects No. 1, No. 28 and No. 32 before and after using the sample is shown in Figure 1 .

[0095] Table 14 Descriptive statistical results of skin moisture content (n=34)

[0096] Statistical analysis results: “-”: no statistically significant difference (P ≥ 0.05); “*”: statistically significant difference (0.01 ≤ P < 0.05); “**”: statistically significant difference (0.001 ≤ P < 0.01); “***”: statistically significant difference (P < 0.001).

[0097] Table 15 Descriptive statistical results of skin elasticity R2 (n=34)

[0098] Statistical analysis results: “***”: statistically significant difference (P < 0.001). P

[0099] Table 16 Descriptive statistical results of skin elasticity R5 (n=34)

[0100] Statistical analysis results: “***”: statistically significant difference (P < 0.001). P

[0101] Table 17 Descriptive statistical results of skin elasticity R7 (n=34)

[0102] Statistical analysis results: “***”: statistically significant difference (P < 0.001).​​P <0.001).

[0103] Table 18 Descriptive statistics of dermal thickness (n=34)

[0104] Statistical analysis results: "-": no statistically significant difference ( P ≥0.05); "*": The difference is statistically significant (0.01≤ P <0.05).

[0105] Table 19 Descriptive statistics of dermal density (n=34)

[0106] Statistical analysis results: "-": no statistically significant difference ( P ≥0.05); "*": The difference is statistically significant (0.01≤ P <0.05).

[0107] As shown in Tables 14-19, during the 4-week test period, the moisturizing and antioxidant face cream, namely Example 4, significantly improved skin moisture content, skin elasticity, dermal thickness and density. Therefore, the face cream product containing this moisturizing and antioxidant composition has good moisturizing, anti-aging and skin elasticity-enhancing effects.

[0108] The above embodiments are only for illustrating the technical concept and features of the present invention, and are intended to enable those skilled in the art to understand the content of the present invention and implement it accordingly. They should not be construed as limiting the scope of protection of the present invention. All equivalent changes or modifications made in accordance with the spirit and essence of the present invention should be covered within the scope of protection of the present invention.

Claims

1. A moisturizing antioxidant composition characterized in that, The moisturizing antioxidant composition comprises the following components in mass fraction: 0.05-1.0 parts of HA-hexapeptide-9 derivative, 1.0-5.0 parts of fritillaria cirrhosa bulb extract, and 1.0-10.0 parts of semen cuscutae extract.

2. A moisturizing antioxidant composition according to Claim 1 wherein, The moisturizing antioxidant composition comprises the following components in mass fraction: 0.1-0.7 parts of HA-hexapeptide-9 derivative, 1.5-4.0 parts of fritillaria cirrhosa bulb extract, and 5.0-8.0 parts of semen cuscutae extract.

3. The moisturizing and antioxidant composition as described in claim 1, characterized in that, The structure of the HA-hexapeptide-9 derivative is as follows: Formula (I): ; wherein .

4. A moisturizing antioxidant composition according to Claim 1 wherein, The preparation method of the fritillaria cirrhosa bulb extract is as follows: dry powder of fritillaria cirrhosa bulb is added into 65-80% ethanol solution, and extraction is carried out for 2-4 times under water bath heating at 60-80°C; the filtrates are combined and concentrated under reduced pressure, and then alcohol is volatilized until no alcohol smell is left; 0.7-1.2 times the mass of butanediol is added, and then stirring and uniform mixing are carried out, and then filtration is carried out to obtain the fritillaria cirrhosa bulb extract; the concentration is 0.25-0.40 g / mL in terms of crude drug.

5. A moisturizing antioxidant composition according to Claim 1 wherein, The preparation method of the semen cuscutae extract comprises the following steps: semen cuscutae powder is subjected to ultrasonic-assisted subcritical extraction to obtain an extraction liquid, and then filtration and concentration are carried out, and then glycerol is added to obtain the semen cuscutae extract.

6. A moisturizing antioxidant composition according to claim 5, wherein the antioxidant is a mixture of ascorbic acid and ascorbyl palmitate. The preparation method of the semen cuscutae extract is as follows: S1: semen cuscutae powder is added into water and subjected to ultrasonic treatment, wherein the mass ratio of material to liquid is 1:5-1:10, the power of ultrasonic waves is set to 300-500 W, the temperature is 25-40°C, and the time is 25-40 min; S2: the material liquid prepared in step S1 is placed in an extractor, 0.5-1.0 times the mass of water of the material liquid prepared in S1 is added, the extraction conditions are as follows: the temperature is 120-140°C, the pressure is 0.1-0.8 MPa, and the time is 30-60 min; the extraction liquid is filtered, the filtrate is concentrated under reduced pressure, 1-1.5 times the mass of glycerol of the concentrated liquid is added, uniform stirring and mixing are carried out, and then filtration is carried out to obtain the semen cuscutae extract; the concentration of the extraction liquid is 0.30-0.45 g / mL in terms of crude drug.

7. A moisturizing antioxidant composition according to any of claims 1 to 6, wherein HA-hexapeptide-9, fritillaria cirrhosa bulb extract and semen cuscutae extract are mixed and dissolved in purified water to make up 100 parts.

8. Use of the moisturizing antioxidant composition according to any one of claims 1 to 6 for antioxidation and moisturization, characterized in that, The antioxidant and moisturizing effects are achieved by improving the expression of GSH, SOD, UCA and PCA.

9. Use of the moisturizing antioxidant composition according to any one of claims 1 to 6 for the preparation of a cosmetic product, characterized in that, The cosmetic has the effects of antioxidant, anti-aging, improvement of facial wrinkles and skin moisture content.

10. A cosmetic product having an antioxidative, anti-aging, wrinkle improvement and skin moisture content-improving effect, characterized by, The cosmetic at least comprises the moisturizing antioxidant composition of claim 7.

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