Anti-aging repairing composition as well as preparation method and application thereof

Abstract

The invention discloses an anti-aging repairing composition as well as a preparation method and application thereof, and particularly belongs to the field of preparations for dressing and makeup. The composition is prepared from the following components: a dendrobium officinale extract, oligopeptide-215, a kangaroo claw extract, an euglena parvata extract and a cymbidium hookerianum extract. The composition provided by the invention has an obvious inhibition effect on inflammatory factors, effectively promotes migration of skin keratinocytes to achieve the effect of repairing the skin, and meanwhile, the composition can promote generation of collagen and elastin fibers to achieve the effects of resisting wrinkles and tightening the skin; the skin external preparation containing the composition provided by the invention has the effects of tightening, resisting wrinkles and repairing.

Description

Technical Field

[0001] The invention belongs to the field of cosmetic preparations, and specifically relates to an anti-aging repair composition and a preparation method and application thereof. Background Art

[0002] With the accelerated pace of life in modern society and the increasing severity of environmental pollution, the problem of skin aging has attracted more and more attention. Skin aging is not only manifested in external changes such as sagging skin and increased wrinkles, but also involves internal changes such as weakened skin barrier function and decreased cell vitality. Therefore, it is particularly important to develop a composition that can effectively resist aging and repair damaged skin.

[0003] Dendrobium officinale extract is rich in polysaccharides, flavonoids, phenols, amino acids and other active ingredients. Studies have shown that these ingredients can effectively remove oxygen free radicals and hydroxyl free radicals, and have antioxidant and anti-aging effects. At the same time, Dendrobium officinale extract also has multiple functions such as antibacterial, anti-inflammatory, and moisturizing, providing a natural and effective choice for anti-aging and skin repair. Oligopeptide-215 is an emerging anti-aging ingredient that has the effect of firming and repairing the skin. It achieves the purpose of anti-aging and skin repair by promoting collagen production, reducing inflammatory expression, and stabilizing the stratum corneum. Kangaroo paw extract is a multi-targeted anti-aging ingredient that can increase the production of tenascin-X, stimulate collagen synthesis, and increase elastin synthesis, thereby improving skin firmness and reducing sagging and wrinkles. Euglena microcarpa extract is mainly used as a skin conditioner in cosmetics. It can soothe and condition the skin, gently remove old dead skin cells, and glow with translucent radiance. It plays an important role in improving skin condition and enhancing skin texture.

[0004] Although the above ingredients have shown good effects in anti-aging and skin repair, the technology of effectively combining these ingredients and applying them to anti-aging and repairing compositions is still imperfect in the prior art. Therefore, it is of great practical significance and application value to develop a composition that can give full play to the synergistic effect of each ingredient, effectively anti-aging and repair damaged skin. Summary of the invention

[0005] In view of the deficiencies of the prior art, the present invention aims to provide a composition in which the components cooperate with each other, effectively resist aging and repair damaged skin.

[0006] To achieve the above object, the present invention discloses the following technical solutions:

[0007] In a first aspect, the present invention provides an anti-aging repair composition, comprising the following components in parts by mass:

[0008] 0.2-1 part of Dendrobium officinale extract;

[0009] Oligopeptide-215 3.8-6.5 parts;

[0010] Kangaroo paw extract 6.7-9.2 parts;

[0011] 10-15 parts of Euglena gracilis extract;

[0012] 1.2-3.0 parts of Cynarostemon fasciatum root extract.

[0013] Dendrobium officinale extract is rich in antioxidant ingredients, which can effectively resist free radical damage to the skin, slow down skin aging, and reduce the appearance of wrinkles and fine lines. At the same time, Dendrobium officinale extract also has excellent moisturizing ability, which can penetrate deep into the skin to provide sufficient moisture and nutrients, keeping the skin supple and smooth. In addition, it can also promote the regeneration and repair of skin cells, relieve skin sensitivity and discomfort, and has anti-inflammatory and antibacterial effects, which helps to improve skin problems such as acne and blackheads.

[0014] As a skin protectant, oligopeptide-215 has significant effects in firming, repairing and promoting collagen. Due to its small molecular weight, it is more easily absorbed by human skin and has good skin permeability.

[0015] Kangaroo paw extract can stimulate collagen synthesis, enhance elastin synthesis, and increase the contractility of fibroblasts, thereby reducing skin sagging, quickly tightening, quickly reducing wrinkles in the eye area, reducing wrinkles in the overall neck area, reducing the depth and length of neck wrinkles, and reducing the roughness of the neck skin.

[0016] Euglena microcarpa extract has good moisturizing ability, which can greatly improve dry skin, prevent water loss, and keep the skin supple and smooth. At the same time, it contains a large amount of polyphenols, which can effectively resist oxidation, prevent excessive production of free radicals, and reduce the speed of skin aging.

[0017] The root extract of Cynanchum paniculatum contains a variety of effective ingredients such as alkaloids, lipids and vitamins, which have the effects of improving skin repair ability, promoting cell proliferation and repair, and stabilizing skin structure to prolong skin aging; the present invention found that the combination of Cynanchum paniculatum root extract with Dendrobium officinale extract, oligopeptide-215 and kangaroo paw extract has a certain promoting effect on the anti-inflammatory and soothing effect of Dendrobium officinale extract, and at the same time has a certain enhancing effect on the performance of Kangaroo paw extract and oligopeptide-215 in promoting the expression of collagen and elastin.

[0018] Preferably, the method for preparing the Cynaropidium root extract comprises the following steps:

[0019] S1. Take dried Cynanchum root, crush and sieve to obtain Cynanchum root powder;

[0020] S2. The Cynanchum root powder obtained in S1 is added to distilled water, and the composite enzyme preparation is added and stirred evenly. After enzymolysis for 2-3 hours, ultrasonic treatment is performed, and the filtrate A and the filter residue are filtered to be used, wherein the composite enzyme preparation includes neutral protease, β-glucanase and mannanase;

[0021] S3. Take an ethanol aqueous solution and add it to the residue obtained in S2, reflux extraction, and filter to obtain a filtrate B;

[0022] S4. After combining filtrates A and B, the filtrates are concentrated and vacuum dried to obtain a Cynarostegium wilfordii root extract.

[0023] Further preferably, in the step S1, the dried Cynanchum root is crushed and then passed through a sieve of ≥60 mesh.

[0024] Further preferably, in step S2, the mass ratio of Cynarostegium wilfordii root powder to distilled water is 1:(20-30);

[0025] The added amount of the complex enzyme preparation is 4-6% of the mass of the Cyperus rotundus root powder;

[0026] In the composite enzyme preparation, the neutral protease activity is ≥50 U / g, the β-glucanase activity is ≥120 U / g, and the mannanase activity is ≥100 U / g.

[0027] Further preferably, in step S2, the ultrasonic temperature is 25-30° C., the ultrasonic frequency is 20-30 kHz, and the ultrasonic time is 20-30 min.

[0028] Further preferably, in step S3, the concentration of the ethanol aqueous solution is ≥95 v / v%, the temperature of the reflux extraction is 85-90° C., and the number of refluxes is 2-3 times, each time for 0.5-0.7 h.

[0029] In a second aspect, the present invention provides a leave-on skin topical preparation with anti-aging and repairing effects, comprising the anti-aging and repairing composition described in the first aspect;

[0030] The amount of the anti-aging repair composition added to the leave-on skin external preparation is 0.01-0.02 wt %.

[0031] Preferably, it also includes a moisturizer, a thickener, a chelating agent, a base oil, an emulsifier, a preservative, a pH adjuster and a solvent;

[0032] The moisturizing agent is at least one of glycerin, butylene glycol, sodium hyaluronate, 1,3-propylene glycol, and 1,2-hexanediol;

[0033] The base oil is at least one of polydimethylsiloxane and hydrogenated palm kernel oil;

[0034] The emulsifier is at least one of cetearyl alcohol and polyglyceryl-6 distearate;

[0035] The thickener is carbomer;

[0036] The chelating agent is disodium EDTA;

[0037] The preservative is p-hydroxyacetophenone;

[0038] The pH regulator is arginine;

[0039] The solvent is deionized water.

[0040] In the third aspect, the present invention provides a method for preparing the leave-on skin topical preparation with anti-aging and repairing effects as described in the second aspect, characterized in that it comprises the following steps: stirring and mixing the moisturizer, thickener, chelating agent, base oil, emulsifier, preservative and solvent at 60-90°C, then cooling to below 45°C, adding the anti-aging and repairing composition and continuing to stir, and finally adding a pH adjuster to adjust the pH to 5.0-6.5 to obtain the leave-on skin topical preparation.

[0041] In a fourth aspect, the present invention provides the use of the anti-aging and repairing composition described in the first aspect in the preparation of an essence, lotion, cream, spray, gel and / or freeze-dried powder with anti-aging and repairing efficacy.

[0042] Beneficial effects of the present invention:

[0043] 1. The anti-aging repair composition provided by the present invention has a significant inhibitory effect on the inflammatory factor TNF-α;

[0044] 2. The anti-aging repair composition provided by the present invention can significantly promote the migration of skin keratinocytes and achieve the effect of repairing the skin;

[0045] 3. The anti-aging repair composition provided by the present invention can promote the expression of COL-I gene in HaCaT human keratinocytes, achieving the effect of anti-wrinkle and firming;

[0046] 4. The anti-aging repair composition provided by the present invention can inhibit the expression of MMP-1 mRNA, slow down the destruction of collagen fibers and elastic fibers by MMP-1, and slow down the aging of the skin;

[0047] 5. The skin topical preparation provided by the present invention has a firming and anti-wrinkle effect, and at the same time, its lactic acid stinging improvement rate is high, and it has a good repair effect. DETAILED DESCRIPTION

[0048] To better illustrate the purpose, technical solutions and advantages of the present invention, the present invention will be further described below in conjunction with specific embodiments. It should be understood by those skilled in the art that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the present invention.

[0049] Unless otherwise specified, the test methods used in the examples and comparative examples are all conventional methods; the materials, reagents, etc. used, unless otherwise specified, can be obtained from commercial channels.

[0050] The extract of Dendrobium officinale is an extract of Dendrobium officinale (DENDROBIUM OFFICINALE), INCI name / English name is DENDROBIUM OFFICINALE EXTRACT, purchased from Fufeng Snort Biotechnology Co., Ltd., specification 20:1;

[0051] Oligopeptide-215 was purchased from Shenzhen Weiqi Technology Co., Ltd.;

[0052] Kangaroo paw extract is kangaroo paw (ANIGOZANTHOS FLAVIDUS) flower extract, INCI name / English name is ANIGOZANTHOS FLAVIDUS EXTRACT, purchased from Shaanxi Aibony Biotechnology Co., Ltd., specification 10:1;

[0053] The Euglena gracilis extract is an extract of Euglena gracilis, INCI name / English name is EUGLENA GRACILIS EXTRACT, purchased from Fufeng Snort Biotechnology Co., Ltd., specification 10:1;

[0054] The root of Cymbidium hookerianum Rchb.F. is a plant of the genus Cymbidium of the family Orchidaceae. In a specific embodiment, dried Cymbidium hookerianum root is used for extraction. The Cymbidium hookerianum root is purchased from Shanghai Zhenrentang Pharmaceutical Co., Ltd.;

[0055] Other raw materials are commercially available.

[0056] Preparation of Cynaraceae Root Extract

[0057] Step S1, taking the dried Cynanchum ovata root and placing it in a high-speed grinder for grinding, and passing it through a ≥60 mesh sieve to obtain Cynanchum ovata root powder;

[0058] Step S2, adding the Cynanchum root powder obtained in S1 to distilled water at room temperature, adding a composite enzyme preparation and stirring evenly, performing ultrasonic treatment after enzymolysis for 2-3 hours, filtering to obtain filtrate A and filter residue for standby use, wherein the mass ratio of the Cynanchum root powder to distilled water is 1:(20-30), the composite enzyme preparation consists of neutral protease, β-glucanase, and mannanase, the neutral protease activity in the composite enzyme preparation is ≥50U / g, the β-glucanase activity is ≥120U / g, and the mannanase is ≥100U / g, the addition amount of the composite enzyme preparation is 4-6% of the mass of the Cynanchum root powder, the ultrasonic temperature is 25-30°C, the ultrasonic frequency is 20-30kHz, and the ultrasonic time is 20-30min;

[0059] Step S3, adding ethanol aqueous solution to the filter residue obtained in S2, performing reflux extraction, and filtering to obtain filtrate B, wherein the concentration of ethanol aqueous solution is 95 v / v%, the amount of ethanol aqueous solution added is 5-10 times the mass of the filter residue, the temperature of reflux extraction is 85-90° C., the number of reflux is 2-3 times, each time for 0.5-0.7 h;

[0060] Step S4, after combining the filtrate A and B, perform reduced pressure distillation and concentrate to 20% of the original volume, and then vacuum freeze-dry for 12-24 hours at a vacuum degree of 5-10 Pa and a temperature of -20--40°C to obtain the Cyperus rotundus root extract. The specific preparation parameters of the Cyperus rotundus root extract are described in Table 1.

[0061] Table 1 Preparation parameters of Cynarostegium wilfordii root extract

[0062]

[0063]

[0064] Note: “-” in the table indicates no addition and related operations.

[0065] Preparation of Examples and Comparative Examples

[0066] Weigh the raw materials according to the mass fractions in Table 2, mix and stir evenly to obtain a composition.

[0067] Table 2 Composition raw materials and their mass proportions

[0068]

[0069] Note: “-” in the table means not added.

[0070] Performance Test:

[0071] Inflammatory factor inhibition experiment

[0072] Samples to be tested: Take a certain amount of the composition of Examples 1-4 and Comparative Examples 1-10, and dilute them to 0.02 wt % with physiological saline.

[0073] Experimental method: RAW264.7 macrophages were used as the research object, and the cell inflammation model was established by stimulating the cells with lipopolysaccharide LPS (bacterial endotoxin). Macrophages were inoculated into 12-well plates and incubated in an incubator at 37°C and 5% CO2 for 24 hours. The dilutions of Examples 1-4 and Comparative Examples 1-10 were added respectively. LPS (1 μg / mL) was added 2 hours later, which was recorded as T1. A group without adding dilution but only adding LPS was set as T 总 The group without LPS and only with diluent was recorded as T0. After 24 hours of stimulation, the supernatant was collected, centrifuged, and tested. The ELISA kit was used to analyze the release level of the pro-inflammatory inflammatory factor TNF-α of RAW264.7. The TNF-α inhibition rate was calculated by the following formula:

[0074] TNF-α inhibition rate (%) = [(T 总 -T1) / (T 总 -T0)]×100%.

[0075] The higher the TNF-α inhibition rate, the better the anti-inflammatory effect of the composition. The results of the TNF-α inhibition test are shown in Table 3.

[0076] Table 3 TNF-α inhibition rate

[0077]

[0078] Result analysis:

[0079] Compared with Comparative Examples 1-10, Examples 1-4 can determine that the composition provided by the present invention has a significant inhibitory effect on the inflammatory factor TNF-α; wherein: compared with Comparative Examples 1-4, the lack of Dendrobium extract, oligopeptide-215, kangaroo paw extract and Stellera root extract in the composition of Example 1 has a greater impact on the anti-inflammatory properties of the composition; the experiment verified that the Dendrobium extract, oligopeptide-215, kangaroo paw extract, microalgae extract and Stellera root extract in the composition have the effect of promoting each other and gaining benefit together.

[0080] Experiment on promoting the healing of skin keratinocyte scratches

[0081] Experimental principle: Cell migration ability directly reflects the activity level of cells. If cells can proliferate and migrate quickly, they can form a protective mechanism as soon as possible in response to exogenous stimuli, thereby repairing damaged cells. The cell scratch test is a simple and quick method to measure cell migration and repair ability, similar to an in vitro wound healing model. Detecting cell migration rate can reflect the barrier repair ability of cells. A large migration rate means that the barrier repair ability of the sample to be tested is good, and a small migration rate means that the barrier repair ability of the sample to be tested is poor.

[0082] Test sample: Take a certain amount of the composition of Examples 1-4 and Comparative Examples 1-10, dilute with physiological saline to a concentration of 0.02 wt%. Blank control: physiological saline. Set three parallel controls for each test sample, and take the average value of the three parallel control results.

[0083] Experimental protocol: HaCat cells were cultured at 4×10 4 / well were inoculated in the cell insert and cultured in a cell culture incubator. When the cell density was about 80%, the cell insert and the liquid in the 6-well plate were removed, 2 ml of each test sample was added, and they were cultured together for 24 hours. Pictures were taken at 0 hour and 24 hours, and the cell area was calculated using ImageJ. The corresponding migration rate was calculated to obtain the effect of the test sample on the migration of skin keratinocytes.

[0084] The calculation formula for the results is: mobility = (0h area - 24h area) / 0h area, relative mobility = sample mobility - blank group mobility.

[0085] The test results are shown in Table 4.

[0086] Table 4 Effects of Examples 1-4 and Comparative Examples 1-10 on the Migration Rate of Skin Keratinocytes

[0087]

[0088] Result analysis:

[0089] The compositions of Examples 1-4 can significantly promote the migration of skin keratinocytes; it can be seen from Example 1 compared with Comparative Examples 1-4 that when the composition lacks any one of oligopeptide-215, kangaroo paw extract, and Stellera fasciata root extract, the repair performance of the composition is greatly reduced. According to the experimental results, it can be seen that Stellera fasciata root extract has a certain skin repair effect, and can promote and enhance the repair effect of oligopeptide-215 and kangaroo paw extract on the skin.

[0090] Collagen COL-Ⅰ mRNA expression level experiment

[0091] Experimental principle: Aging is a complex process, and wrinkles are an important evidence and external manifestation of aging. The dermis is closely related to the formation of wrinkles and is mainly responsible for the production of collagen and elastin fibers. Collagen fibers form a dense mesh structure to maintain skin cell organization and resistance, while finer elastin fibers make the skin soft and elastic. The number of collagen fibers and elastic fibers decreases with skin aging. The reduction of dermal collagen fibers, the disorder of collagen fiber bundle structure, and the activation or increase of MMPs caused by ultraviolet rays will lead to the decomposition and breakage of collagen, thereby causing the skin to lose elasticity and form wrinkles. Therefore, the expression level of collagen genes can reflect the collagen content of the skin, and then directly reflect whether the composition has anti-wrinkle ability, thereby reflecting its anti-aging effect.

[0092] Samples: (1) Control group: DMEM culture medium containing 10% serum; (2) Test sample group: The compositions of Examples 1-4 and Comparative Examples 1-10, prepared into a solution with a concentration of 0.02 wt % using DMEM culture medium containing 10% serum as solvent.

[0093] Experimental steps: HaCaT human keratinocytes were seeded in 6-well plates (2×10 5 Each well of the test sample group was added with the test sample solution and cultured for 24 hours. After the culture was completed, the supernatant was discarded and 30 mJ / cm 2 HaCaT human keratinocytes were irradiated with UVB for 30 min, while the control group was cultured normally without UV irradiation. The expression of collagen-related gene COL-Ⅰ was detected by qRT-PCR.

[0094] Expression of experimental results: The higher the expression level of COL-Ⅰ gene, the better the anti-wrinkle ability.

[0095] The experimental results are shown in Table 5:

[0096] Table 5 COL-Ⅰ mRNA expression

[0097]

[0098]

[0099] The test results show that the compositions of the embodiments and comparative examples of the present application can promote the expression of COL-I gene in HaCaT human keratinocytes, indicating that the compositions of the embodiments and comparative examples of the present application have anti-wrinkle ability. Among them, the anti-wrinkle effect of Examples 1-4 is better than that of Comparative Examples 1-4, indicating that in the present application, the combination of Dendrobium extract, oligopeptide-215, kangaroo paw extract and Stellera root extract can promote the anti-wrinkle effect of the composition. Without any of the components, the anti-wrinkle effect of the composition is poor. Comparison of Examples 1-4 with Comparative Example 10 shows that the ratio of Dendrobium extract, oligopeptide-215, kangaroo paw extract and Stellera root extract will also affect the overall anti-wrinkle effect of the composition.

[0100] MMP-1 mRNA expression level experiment

[0101] Experimental principle: One of the main causes of skin aging is the change in the structure of the dermis, and there are many reasons for the structural change. External factors activate matrix metalloproteinases (MMPs) in the body, which will lead to excessive degradation of collagen and elastin in the dermis that support the skin structure, resulting in wrinkles, decreased elasticity and other aging symptoms. The degradation of the extracellular matrix mainly depends on proteolytic enzymes, and MMPs are the most important group of proteolytic enzymes, among which MMP-1 is the most important enzyme for degrading type I and type III collagen. When the mRNA of MMP-1 is overexpressed, it indicates that the content of MMPs is high. MMPs will specifically degrade the components of the extracellular matrix and destroy the normal structure of collagen fibers and elastic fibers, resulting in wrinkles and other aging manifestations of the skin.

[0102] MMP-1mRNA test method:

[0103] Samples: (1) Control group: DMEM culture medium containing 10% serum; (2) Test sample group: The compositions of Examples 1-4 and Comparative Examples 1-10, prepared into a solution with a concentration of 0.02 wt % using DMEM culture medium containing 10% serum as solvent.

[0104] Experimental steps: HaCaT human keratinocytes were seeded in 6-well plates (2×10 5 Each well of the sample group was added with the sample solution and cultured for 24 hours. After the culture was completed, the supernatant was discarded and 30 mJ / cm 2 HaCaT human keratinocytes were irradiated with ultraviolet B (UVB) for 1 h. The expression of collagen-related gene MMP-1 was detected by qRT-PCR.

[0105] Detection index: The higher the expression level of MMP-1 mRNA, the more MMP-1 there is, the easier it is for the collagen fibers and elastic fibers in the skin to be damaged, and the easier it is for the skin to have wrinkles and aging problems. The results are shown in Table 6.

[0106] Table 6 MMP-1 mRNA expression

[0107] Group MMP-1 mRNA expression Control group 4.12 Example 1 1.98 Example 2 2.00 Example 3 1.94 Example 4 1.92 Comparative Example 1 2.97 Comparative Example 2 3.01 Comparative Example 3 3.03 Comparative Example 4 2.89 Comparative Example 5 3.91 Comparative Example 6 3.44 Comparative Example 7 3.39 Comparative Example 8 3.42 Comparative Example 9 3.53 Comparative Example 10 2.27

[0108] Result analysis:

[0109] Compared with the control group, the compositions of the examples of the present application and the comparative examples can inhibit the expression of MMP-1 mRNA, slow down the destruction of collagen fibers and elastic fibers by MMP-1, and slow down skin aging.

[0110] According to the comparison between Example 1 and Comparative Examples 1-4, the expression level of MMP-1 mRNA in Example 1 is lower than that in Comparative Examples 1-4, indicating that in the composition of the present application, there is a synergistic effect between the Dendrobium extract, oligopeptide-215, kangaroo paw extract and Cynaraceae root extract, and the lack of one of the components will result in a poorer inhibitory effect of the composition on MMP-1 mRNA.

[0111] Preparation of application examples

[0112] The specific components are weighed according to the mass percentage of raw materials in Table 7.

[0113] Step (1), mixing component A with water, heating to 90° C., homogenizing at a speed of 1200 rpm for 10 min, and keeping warm for later use after homogenization to obtain prefabricated component A;

[0114] Step (2), mixing component B, heating to 85° C., homogenizing at a speed of 1200 rpm for 10 min, and keeping warm for later use after homogenization to obtain prefabricated component B;

[0115] Step (3) Mix the C components, heat to 75° C., melt and stir evenly to obtain a prefabricated C component;

[0116] Step (4) The preformed component A is heated to 80°C, and the preformed component B is added at a speed of 300 rpm, and stirred and mixed. Then, the temperature is lowered to 60°C, and the preformed component C is added at a speed of 300 rpm and stirred and mixed. Then, the temperature is lowered to below 45°C, and the composition of Example 1 and the essence are added and stirring is continued for 5 minutes. Finally, arginine is added to adjust the pH to 6.5, and stirring can be stopped. The material is sieved through a 200-mesh sieve to obtain a residual skin topical preparation.

[0117] Table 7: Raw materials of leave-on skin topical preparations by weight (wt%)

[0118]

[0119] Application performance test

[0120] Multiple skin irritation tests:

[0121] Materials and methods:

[0122] 1. Test substance: Use the product of Application Examples 1-4 directly.

[0123] 2. Experimental animals and breeding environment

[0124] 2.1 Experimental animals: 4 female Japanese large-eared white rabbits, weighing 2.0kg-2.5kg, provided by Beijing Changyang Xishan Breeding Farm, ordinary grade animals.

[0125] 2.2 Animal room temperature was controlled at 19-26°C, relative humidity was 40-70%, lighting was 12 hours, darkness was 12 hours. The cages were stainless steel, and the animals were raised in single cages. Animals were allowed to eat and drink freely.

[0126] 3. Test methods

[0127] 3.1 24 hours before the test, remove the hair on both sides of the rabbit's spine, about 3cm×3cm on each side, without damaging the skin.

[0128] 3.2 Apply 0.5 ml of the test substance directly to the skin of both sides of the animal (2.5 cm × 2.5 cm) once a day for 14 consecutive days. From the second day, remove the hair before application and use water to remove the residual test substance. Observe the results after one hour.

[0129] 3.3 Result evaluation: Skin irritation reaction scores were performed according to the relevant provisions of the "Technical Specifications for Safety of Cosmetics" (2015 edition), and the average score of each animal per day was calculated. The average score of each animal per day = Σ erythema and edema score / number of test animals / 14, and the skin irritation intensity was graded. The results are shown in Table 8.

[0130] Table 8

[0131]

[0132] Note: The integral mean is rounded to 2 decimal places.

[0133] Result analysis:

[0134] The test substance was used to conduct multiple skin irritation test on domestic rabbits. The average score for each animal per day was 0.00 (0~<0.5). The test substance was non-irritating to the multiple skin irritation of domestic rabbits.

[0135] Human skin patch test:

[0136] The four application examples prepared above were subjected to the human skin patch test in the "Technical Specification for Safety of Cosmetics" (2015 edition).

[0137] The skin reaction was observed according to the standard at 30 minutes (after the indentation disappears), 24 hours and 48 hours, and the observation results were recorded. See Table 9.

[0138] Table 9 Human safety test results

[0139]

[0140]

[0141] The results of human skin patch tests showed that no adverse skin reactions occurred among 30 people.

[0142] Firming and anti-wrinkle human efficacy test

[0143] Experimental method: 25 people with facial wrinkles (30-45 years old) were selected as volunteers for the firming and anti-wrinkle efficacy test (5 volunteers per group, each group used the same test sample, and the control group used deionized water). The volunteers used the test sample on their entire face, about 2 mL / time, once in the morning and evening every day, and measured the change rate of their skin firmness R0 and skin elasticity R2 in the 0th and 2nd week of use, respectively, to evaluate the effect of resident skin topical agents in reducing skin wrinkles.

[0144] Test instrument: Skin elasticity tester MPA580 (produced by CK Company of Germany).

[0145] The higher the skin firmness R0 value, the firmer the skin is; the higher the skin elasticity R2 value, the better the skin elasticity is.

[0146] Skin firmness R0 change rate = (skin firmness R0 value after 2 weeks of use - skin firmness R0 value before use) / skin firmness R0 value before use × 100%.

[0147] Skin elasticity R2 change rate = (skin elasticity R2 value after 2 weeks of use - skin elasticity R2 value before use) / skin elasticity R2 value before use × 100%.

[0148] The test results are shown in Table 10.

[0149] Table 10 Firming and anti-wrinkle performance test results

[0150] Test samples Skin firmness R0 change rate (%) Skin elasticity R2 change rate (%) Control group -1.46 -0.81 Application Example 1 20.36 18.91 Application Example 2 19.47 18.22 Application Example 3 17.28 17.43 Application Example 4 6.25 5.16

[0151] Repair performance test

[0152] Volunteer screening: 25 people with sensitive skin (25-45 years old) were selected as volunteers for the repair performance test (every 5 volunteers used the same test sample).

[0153] Test steps: The test was conducted on the nasolabial folds of volunteers. 50ul of 10% lactic acid solution was dropped on a single-layer filter paper with a diameter of 0.8cm. The subjects were asked about the tingling sensation at 0.5min and 2min respectively. The pain was scored according to the 4-point system (0 for no tingling, 1 for mild tingling, 2 for moderate tingling, and 3 for severe tingling). If the subject reached 3 points at any time during the test, the test was terminated.

[0154] Sample use: Volunteers used the leave-on skin topical preparations of Examples 1-4 on their entire faces, and volunteers in the blank control group used deionized water as samples. Volunteers applied the product twice a day, morning and evening, about 1 mL, for two consecutive weeks, and repeated the above test steps after two weeks of use.

[0155] The lactic acid stinging scores of the volunteers before use and after 2 weeks of use were recorded, and the lactic acid stinging improvement rate of the volunteers was calculated. Wherein, lactic acid stinging improvement rate (%) = (average lactic acid stinging score before use - average lactic acid stinging score after 2 weeks of use) / average lactic acid stinging score before use × 100%; average lactic acid stinging score = average stinging score at 0.5 min + average stinging score at 2 min.

[0156] The higher the improvement rate of lactic acid stinging, the better the repair effect of the sample. The results are shown in Table 11.

[0157] Table 11 Lactic acid stinging improvement rate

[0158]

[0159] Result analysis:

[0160] The test results in Tables 10 and 11 show that the leave-on skin topical preparations of Examples 1-3 have excellent firming and anti-wrinkle effects. At the same time, the skin topical preparations have a high improvement rate of lactic acid stinging and have a good repairing effect.

[0161] Finally, it should be noted that the above embodiments and comparative examples are only used to illustrate the technical solution of the present invention rather than to limit the protection scope of the present invention. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solution of the present invention can be modified or replaced by equivalents without departing from the essence and scope of the technical solution of the present invention.

Claims

1. An anti-aging repair composition, characterized in that: By mass, it includes the following components: 0.2-1 part of Dendrobium officinale extract; Oligopeptide-215 3.8-6.5 parts; Kangaroo paw extract 6.7-9.2 parts; 10-15 parts of Euglena gracilis extract; 1.2-3.0 parts of Cynarostemon fasciatum root extract.

2. The anti-aging repair composition according to claim 1, characterized in that: The preparation method of the Cynaraceae root extract comprises the following steps: S1. Take dried Cynanchum root, crush and sieve to obtain Cynanchum root powder; S2. The Cynanchum root powder obtained in S1 is added to distilled water, and the composite enzyme preparation is added and stirred evenly. After enzymolysis for 2-3 hours, ultrasonic treatment is performed, and the filtrate A and the filter residue are filtered to be used, wherein the composite enzyme preparation includes neutral protease, β-glucanase and mannanase; S3. Take an ethanol aqueous solution and add it to the residue obtained in S2, reflux extraction, and filter to obtain a filtrate B; S4. After combining filtrates A and B, the filtrates are concentrated and vacuum dried to obtain a Cynarostegium wilfordii root extract.

3. The anti-aging repair composition according to claim 2, characterized in that: In the step S1, the dried Cynanchum root is crushed and then passed through a sieve with mesh size of ≥60.

4. The anti-aging repair composition according to claim 2, characterized in that: In step S2, the mass ratio of Cynaraceae root powder to distilled water is 1:(20-30); The added amount of the complex enzyme preparation is 4-6% of the mass of the Cynaraceae root powder; In the composite enzyme preparation, the neutral protease activity is ≥50 U / g, the β-glucanase activity is ≥120 U / g, and the mannanase activity is ≥100 U / g.

5. The anti-aging repair composition according to claim 2, characterized in that: In step S2, the ultrasonic temperature is 25-30° C., the ultrasonic frequency is 20-30 kHz, and the ultrasonic time is 20-30 min.

6. The anti-aging repair composition according to claim 2, characterized in that: In step S3, the concentration of the ethanol aqueous solution is ≥95 v / v%, the temperature of the reflux extraction is 85-90° C., and the number of refluxes is 2-3 times, each time for 0.5-0.7 h.

7. A leave-on skin topical preparation with anti-aging and repairing effects, characterized in that: An anti-aging repair composition comprising any one of claims 1 to 6; The amount of the anti-aging repair composition added to the leave-on skin external preparation is 0.01-0.02 wt %.

8. The leave-on skin external preparation with anti-aging and repairing effects according to claim 7, characterized in that: It also includes humectants, thickeners, chelating agents, base oils, emulsifiers, preservatives, pH adjusters and solvents; The moisturizing agent is at least one of glycerin, butylene glycol, sodium hyaluronate, 1,3-propylene glycol, and 1,2-hexanediol; The base oil is at least one of polydimethylsiloxane and hydrogenated palm kernel oil; The emulsifier is at least one of cetearyl alcohol and polyglyceryl-6 distearate.

9. A method for preparing the residual skin external preparation with anti-aging and repairing effects as claimed in claim 8, characterized in that: The following steps are involved: The moisturizer, thickener, chelating agent, base oil, emulsifier, preservative and solvent are stirred and mixed at 60-90° C., then cooled to below 45° C., the anti-aging repair composition is added and continued to be stirred, and finally a pH adjuster is added to adjust the pH to 5.0-6.5 to obtain a leave-on skin topical preparation.

10. Use of the anti-aging and repairing composition according to any one of claims 1 to 6 in the preparation of an essence, lotion, cream, spray, gel and / or freeze-dried powder with anti-aging and repairing effects.

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