Tightening and anti-aging composition containing Campanumoea maculata bark extract and application of firming and anti-aging composition

Abstract

The invention discloses a firming and anti-aging composition containing a Campania acuta bark extract and application of the firming and anti-aging composition, and belongs to the technical field of cosmetics. The firming and anti-aging composition containing the Campanumoea maculata bark extract comprises the following components: the Campanumoea maculata bark extract, a Flos Magnoliae Officinalis extract, a Tremella aurantialba extract and a composite polypeptide, and the composite polypeptide is composed of carnosine, acetyl tetrapeptide-5 and tripeptide-10 citrulline in a mass ratio of 1: (0.4-0.6): (0.2-0.6). Through the synergistic effect of the four components, the firming and anti-aging composition has a good effect of inhibiting the activity of elastase and collagenase.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of cosmetics, in particular to a firming and anti-aging composition containing Erythrina variegate tree bark extract and application thereof. BACKGROUND

[0002] With the aggravation of population aging and the increasing environmental pollution, skin aging has become a global focus. Skin aging is mainly manifested as increased wrinkles, decreased elasticity, and formed color spots, and its core mechanism involves free radical oxidative damage, collagen degradation, and extracellular matrix (ECM) structure destruction. Studies have shown that external factors such as ultraviolet (UV) and air pollutants can induce the generation of a large amount of reactive oxygen species (ROS), activate the mitogen-activated protein kinase (MAPK) signaling pathway, and then up-regulate the expression of matrix metalloproteinases (MMPs, such as collagenase MMP-1 and elastase MMP-9), leading to irreversible degradation of collagen fibers and elastic fibers. In addition, endogenous factors such as cell replicative senescence caused by telomere shortening and fibroblast functional decline further accelerate this process.

[0003] At present, anti-aging cosmetics mainly rely on the following technical paths: (1) antioxidant ingredients (such as vitamin C and coenzyme Q10) reduce oxidative damage by scavenging free radicals; (2) peptide ingredients (such as palmitoyl pentapeptide-4) promote collagen synthesis by activating the TGF-β signaling pathway; (3) plant extracts (such as asiaticoside) reduce MMPs activity by inhibiting the NF-κB pathway. However, single component often has problems such as limited target site, low bioavailability or poor stability. In addition, the existing technology has limited synergistic inhibitory effect on collagenase and elastase, making it difficult to achieve multi-level anti-aging.

[0004] Based on the above problems, there is an urgent need to provide a firming and anti-aging composition that has good inhibitory effect on collagenase and elastase for consumers to choose. SUMMARY

[0005] The purpose of the present application is to provide a firming and anti-aging composition containing Erythrina variegate tree bark extract and application thereof, which uses natural plant extracts and composite polypeptides for compounding, has high safety, and has significant firming and anti-aging effects.

[0006] In order to achieve the above purpose, the present application adopts the following technical scheme: In a first aspect, the present application provides a firming and anti-aging composition containing Erythrina variegate tree bark extract, which comprises the following components: Erythrina variegate tree bark extract, Jasminum nudiflorum flower extract, Auricularia auricula extract, and composite polypeptide.

[0007] Preferably, the firming and anti-aging composition containing Erythrina variegate tree bark extract comprises the following components by mass fraction: Extracfs of Excoecaria acida bark: 1-3 parts; Extracfs of Jasminum nudiflorum: 0.5-0.7 parts; Extracfs of Tremella fuciformis: 2-4 parts; Compound polypeptide: 0.3-0.5 parts: The compound polypeptide comprises at least one of carnosine, acetyl tetrapeptide-5, and tripeptide-10 citrulline.

[0008] Further preferably, the compound polypeptide consists of carnosine, acetyl tetrapeptide-5, and tripeptide-10 citrulline in a mass ratio of 1:0.4-0.6:0.2-0.6.

[0009] In a second aspect, the present application provides use of the firming and anti-aging composition of the first aspect in the preparation of a cosmetic product having firming and anti-aging effects.

[0010] Preferably, the cosmetic product having firming and anti-aging effects is in any one of a cream, a lotion, and an essence.

[0011] In a third aspect, the present application provides a lotion having firming and anti-aging effects, which consists of the following components in mass percentage: The firming and anti-aging composition of the first aspect containing extracfs of Excoecaria acida bark: 1-3 wt%; Humectant: 1-5 wt%; Emollient: 1-5 wt%; Emulsifier: 0.5-3 wt%; Preservative: 0.2-0.5 wt%; Deionized water is supplemented to 100 wt%.

[0012] The humectant is one or more of methylpropanediol, 1,3-propanediol, and butylene glycol; The emollient is one or more of squalane, white pool seed oil, shea butter, European hazel seed oil, and jojoba oil; The preservative is one or more of phenoxyethanol, sodium benzoate, p-hydroxyacetophenone, and sorbic acid; The emulsifier is one or more of C14-22 alcohol, C12-20 alkyl glucoside, and polyglyceryl-3 methylglucose distearate.

[0013] In a fourth aspect, the present application provides a preparation method of the lotion of the third aspect, and the specific steps are as follows: S1: mix the moisturizer, preservative and 1 / 2 deionized water, heat to 60-80℃ and stir, stirring speed: 100-300 rpm, stirring time: 1-3 min; then homogenize, homogenization speed: 3000-5000 rpm, homogenization time: 1-3 min, to obtain a mixed solution A; S2: mix the emollient and emulsifier, heat to 60-80℃ and stir, stirring speed: 200-400 rpm, stirring time: 1-3 min; then homogenize, homogenization speed: 3000-5000 rpm, homogenization time: 1-3 min, to obtain a mixed solution B; S3: add the mixed solution B to the mixed solution A, stir, stirring speed: 200-400 rpm, stirring time: 3-5 min; then homogenize, homogenization speed: 3000-5000 rpm, homogenization time: 3-5 min, to obtain a mixed solution; S4: mix the firming and anti-aging composition and the remaining deionized water, heat to 30-40℃ and stir, stirring speed: 100-300 rpm, stirring time: 1-3 min, to obtain a mixed solution C; S5: after the mixed solution is cooled to 30-50℃, mix the mixed solution C and the mixed solution, stir, stirring speed: 100-300 rpm, stirring time: 1-3 min; then homogenize, homogenization speed: 1000-3000 rpm, homogenization time: 1-3 min, continue to cool to 20-25℃, to obtain a firming and anti-aging emulsion, which is filled, sterilized, sealed and stored at room temperature.

[0014] Compared with the prior art, the present application has the following advantages: The firming and anti-aging composition containing the bark extract of Eucalyptus punctata contains the bark extract of Eucalyptus punctata, the flower extract of Litchi chinensis, the extract of Auricularia auriculajudae and a composite polypeptide with a specific composition (i.e. carnosine, acetyl tetrapeptide-5 and tripeptide-10 citrulline with a mass ratio of 1:0.4-0.6:0.2-0.6), which realizes the synergistic effect of multiple targets and multiple levels.

[0015] Specifically, the bark extract of Eucalyptus punctata is rich in polyphenols and saponins, has strong antioxidant properties, can neutralize free radicals and reduce the expression of matrix metalloproteinases (MMPs, including elastase and collagenase) induced by oxidative stress; part of the polyphenols can directly bind to the enzyme active center to inhibit its catalytic function.

[0016] The flower extract of Litchi chinensis contains magnolin, volatile oil and flavonoids (such as rutin), can inhibit the release of inflammatory factors (such as TNF-α and IL-1β), and the inflammatory factors are key upstream inducers of MMPs transcription; at the same time, flavonoids may directly weakly inhibit the activity of MMPs.

[0017] The main active ingredient of Auricularia auricula extract is Auricularia auricula polysaccharide (such as acidic heteropolysaccharide), which can reduce enzyme activity by chelating metal ions (such as Zn²⁺, Cu²⁺, essential cofactors of MMPs); in addition, the moisturizing effect of polysaccharide can maintain the hydration state of extracellular matrix (ECM), indirectly protecting collagen / elastin.

[0018] Carnosine has both antioxidant (scavenging hydroxyl radicals, capturing carbonyl groups) and metal chelating (binding Cu²⁺ / Zn²⁺) abilities, and can inhibit metal ion-dependent MMPs; at the same time, its anti-glycosylation effect reduces collagen cross-linking damage, indirectly maintaining the enzyme inhibition environment.

[0019] Acetyl tetrapeptide-5 belongs to metalloproteinase inhibitor peptides (such as targeting MMP-1, MMP-9), which may compete with enzyme substrates for binding to the active site, or inhibit the secretion of MMPs (such as reducing the mRNA expression of MMPs in fibroblasts).

[0020] Tripeptide-10 citrulline contains citrulline, which can act as a signal peptide to promote collagen synthesis in fibroblasts; at the same time, the citrulline residue can interact with key residues in the enzyme active center, weakly inhibiting elastase or MMPs.

[0021] The above four components exert synergistic effects through the following pathways: The extract of Malpighia emarginata bark (polyphenols), carnosine (metal chelation), and acetyl tetrapeptide-5 (competitive inhibition) can target the active center, cofactor, or substrate binding site of elastase / MMPs, respectively, forming a "multisite blockade"; the extract of Jasminum mesnyi (anti-inflammatory to reduce factor induction) and acetyl tetrapeptide-5 (inhibiting MMPs transcription) can down-regulate the gene expression of MMPs, reducing enzyme production from the source; the antioxidant effects of the extract of Malpighia emarginata bark (polyphenols) and carnosine can reduce ROS levels, and ROS is an important trigger for MMPs activation (such as through oxidative modification of enzyme precursors or activation of protease-activated receptors), and their synergistic effect not only directly inhibits enzymes but also reduces the chances of their "activation", amplifying the inhibitory effect; carnosine (strong metal chelation) and Auricularia auricula extract (weak chelation) can synergistically reduce the concentration of Zn²⁺ / Cu²⁺ in the local environment, significantly weakening the activity of MMPs (such as MMP-2, MMP-9) that depend on these ions, forming a "1+1>2" chelation effect; tripeptide-10 citrulline (promoting collagen synthesis) and other components (inhibiting enzyme decomposition) form a "synthesis-protection" double path: on the one hand, increasing the total amount of ECM substrates (collagen / elastin), and on the other hand, reducing their degradation, together delaying skin aging.

[0022] The above synergistic mechanism breaks through the limitation of single target of traditional anti-aging ingredients, and significantly improves the synergistic inhibition effect on collagenase and elastase activity. In addition, the scientific proportion of the composition ensures the mildness and safety of the formula, and can be adapted to various dosage forms, providing technical support for the development of high-efficiency synergistic anti-aging cosmetics. DETAILED DESCRIPTION

[0023] In order to better understand the present application, the present application will be further described below in conjunction with specific examples, wherein the terms used in the examples are used to describe specific specific embodiments, and do not constitute a limitation on the scope of protection of the present application.

[0024] The experimental methods in the following tests without specific conditions are usually according to the conventional conditions, or according to the conditions suggested by the manufacturers. Unless otherwise specified, the percentages and parts are calculated by weight.

[0025] Some raw materials and sources are as follows: Plectranthus verticillatus bark: purchased from Shanghai Qianfei Chemical Co., Ltd.

[0026] Jasminum nudiflorum Lindl flower extract: purchased from Shanghai Jia Kai Biological Co., Ltd.

[0027] Gold ear extract: purchased from Shanghai Lanhai Biological Technology Co., Ltd.

[0028] Carnosine: purchased from Shenzhen Ruide Lin Biological Technology Co., Ltd.

[0029] Acetyl tetrapeptide-5: purchased from Shenzhen Ruide Lin Biological Technology Co., Ltd.

[0030] Tripeptide-10 citrulline: purchased from Shenzhen Ruide Lin Biological Technology Co., Ltd.

[0031] Tremella fuciformis extract: purchased from Shanghai Huiwen Biological Technology Co., Ltd.

[0032] Other raw materials or reagents are all conventional market sales.

[0033] The preparation method of the Plectranthus verticillatus bark extract described in the present application is as follows: S1. Dry the Plectranthus verticillatus bark to a water content of ≤5%, and grind it into coarse powder; S2. Mix the Plectranthus verticillatus bark coarse powder with rapeseed oil and polyglyceryl-3 diisostearate, heat to 55℃, and stir at 300 rpm for 20 min to obtain a mixed solution; S3. Under N2 atmosphere, ultrasonic-assisted extraction is performed on the mixed solution in S2 to obtain an extract, the extraction temperature is 55℃, the ultrasonic frequency is 30 kHz, and the extraction time is 45 min; S4. The extract is filtered to remove insoluble substances, centrifuged, and the supernatant is taken to obtain the Euclea natalensis bark extract: The mass ratio of the Euclea natalensis bark coarse powder, rapeseed oil, and polyglyceryl-3 diisostearate in step S2 is 1:3:0.2.

[0034] The mass parts of the components in the firming and anti-aging composition containing the Euclea natalensis bark extract according to the present application are as follows: Firming and anti-aging composition 1: Euclea natalensis bark extract: 2 parts; Jasminum mesnyi flower extract: 0.6 parts; Auricularia auricula extract: 3 parts; and complex polypeptide: 0.4 parts, wherein the complex polypeptide is composed of camosine, acetyl tetrapeptide-5, and tripeptide-10 arginine in a mass ratio of 1:0.5:0.4.

[0035] Firming and anti-aging composition 2: Euclea natalensis bark extract: 3 parts; Jasminum mesnyi flower extract: 0.7 parts; Auricularia auricula extract: 4 parts; and complex polypeptide: 0.3 parts, wherein the complex polypeptide is composed of camosine, acetyl tetrapeptide-5, and tripeptide-10 arginine in a mass ratio of 1:0.6:0.2.

[0036] Firming and anti-aging composition 3: Euclea natalensis bark extract: 1 part; Jasminum mesnyi flower extract: 0.5 parts; Auricularia auricula extract: 2 parts; and complex polypeptide: 0.5 parts, wherein the complex polypeptide is composed of camosine, acetyl tetrapeptide-5, and tripeptide-10 arginine in a mass ratio of 1:0.4:0.6.

[0037] Firming and anti-aging composition ①: Different from composition 1, the Euclea natalensis bark extract is absent, and the absence is supplemented by 0.6 parts of Jasminum mesnyi flower extract, 3 parts of Auricularia auricula extract, and 0.4 parts of complex polypeptide, wherein the complex polypeptide is composed of camosine, acetyl tetrapeptide-5, and tripeptide-10 arginine in a mass ratio of 1:0.5:0.4.

[0038] Firming and anti-aging composition ②: Different from composition 1, the Jasminum mesnyi flower extract is absent, and the absence is supplemented by 2 parts of Euclea natalensis bark extract, 3 parts of Auricularia auricula extract, and 0.4 parts of complex polypeptide, wherein the complex polypeptide is composed of camosine, acetyl tetrapeptide-5, and tripeptide-10 arginine in a mass ratio of 1:0.5:0.4.

[0039] Firming and anti-aging composition ③: The difference between the composition 1 is that it lacks the Auricularia auricula extract, which is replaced by the extract of the tree bark of the tree Eucalyptus globulus Labill. in an amount of 0.6 parts by mass, the extract of the flower of the plant Jasminum sambac in an amount of 3 parts by mass, and the complex polypeptide consisting of the carnosine, the acetyl tetrapeptide-5, and the tripeptide-10-citrulline in a mass ratio of 1:0.5:0.4 in an amount of 2 parts by mass.

[0040] The firming and anti-aging composition ④: The difference between the composition 1 is that it lacks the complex polypeptide, which is replaced by the extract of the tree bark of the tree Eucalyptus globulus Labill. in an amount of 0.6 parts by mass, the extract of the flower of the plant Jasminum sambac in an amount of 3 parts by mass, and the Auricularia auricula extract in an amount of 0.4 parts by mass.

[0041] The firming and anti-aging composition ⑤: The difference between the composition 1 is that the complex polypeptide is replaced by an equal amount of carnosine, and the rest of the raw materials and the amount by mass are the same as those of the composition 1.

[0042] The firming and anti-aging composition ⑥: The difference between the composition 1 is that the complex polypeptide is replaced by an equal amount of acetyl tetrapeptide-5, and the rest of the raw materials and the amount by mass are the same as those of the composition 1.

[0043] The firming and anti-aging composition ⑦: The difference between the composition 1 is that the complex polypeptide is replaced by an equal amount of tripeptide-10-citrulline, and the rest of the raw materials and the amount by mass are the same as those of the composition 1.

[0044] The firming and anti-aging composition ⑧: The difference between the composition 1 is that the complex polypeptide consists of carnosine, acetyl tetrapeptide-5, and tripeptide-10-citrulline in a mass ratio of 1:0.2:0.7, and the rest of the raw materials and the amount by mass are the same as those of the composition 1.

[0045] The firming and anti-aging composition ⑨: The extract of the tree bark of the tree Eucalyptus globulus Labill.: 0.6 parts; the extract of the flower of the plant Jasminum sambac: 3 parts; the Auricularia auricula extract: 0.4 parts; the complex polypeptide: 2 parts; the complex polypeptide consists of carnosine, acetyl tetrapeptide-5, and tripeptide-10-citrulline in a mass ratio of 1:0.5:0.4.

[0046] The firming and anti-aging composition ⑩: The difference between the composition 1 is that it lacks the Auricularia auricula extract, which is replaced by the extract of the tree bark of the tree Eucalyptus globulus Labill. in an amount of 0.6 parts by mass, the extract of the flower of the plant Jasminum sambac in an amount of 3 parts by mass, and the complex polypeptide consisting of the carnosine, the acetyl tetrapeptide-5, and the tripeptide-10-citrulline in a mass ratio of 1:0.5:0.4 in an amount of 2 parts by mass. An emulsion with firming and anti-aging effects, the mass percentage of its components is shown in the following Table 1.

[0047] Table 1 Components of the emulsion with firming and anti-aging effects and their mass percentages

[0048] Note: the humectant is 1,3-propanediol; the emulsifier is C14-22 alcohol, C12-20 alkyl glucoside, polyglyceryl-3 methyl glucose distearate with a mass ratio of 1:1:1; the emollient is jojoba oil, white pool seed oil, European hazel seed oil with a mass ratio of 1:0.5:0.7; the preservative is p-hydroxyacetophenone.

[0049] Blank emulsion: The only difference between the blank emulsion and Emulsion 1 is that the blank emulsion does not contain the firming and anti-aging composition, and the missing mass percentage is made up with deionized water.

[0050] The specific steps of the preparation methods of Emulsions 1-5 and Emulsions ①-⑩ are as follows: S1: Mix the humectant, preservative, and 1 / 2 deionized water, heat to 70°C and stir, stirring speed: 300 rpm, stirring time: 2 min; then homogenize, homogenization speed: 4000 rpm, homogenization time: 2 min, to obtain mixed liquid A; S2: Mix the emollient and emulsifier, heat to 70°C and stir, stirring speed: 300 rpm, stirring time: 2 min; then homogenize, homogenization speed: 4000 rpm, homogenization time: 2 min, to obtain mixed liquid B; S3: Add mixed liquid B to mixed liquid A, stir, stirring speed: 300 rpm, stirring time: 5 min; then homogenize, homogenization speed: 4000 rpm, homogenization time: 5 min, to obtain mixed liquid; S4: Mix the firming and anti-aging composition with the remaining deionized water, heat to 35°C and stir, stirring speed: 300 rpm, stirring time: 2 min, to obtain mixed liquid C.

[0051] S5: After the mixed liquid is cooled to 40°C, mix mixed liquid C with the mixed liquid, stir, stirring speed: 300 rpm, stirring time: 2 min; then homogenize, homogenization speed: 2000 rpm, homogenization time: 2 min, continue to cool to 25°C, to obtain an emulsion with firming and anti-aging effects, fill, sterilize, seal, and store at room temperature.

[0052] Blank emulsion preparation steps: S1: Mix the humectant, preservative, and 1 / 2 deionized water, heat to 70°C and stir, stirring speed: 300 rpm, stirring time: 2 min; then homogenize, homogenization speed: 4000 rpm, homogenization time: 2 min, to obtain mixed liquid A; S2: mix the emollient with the emulsifier, heat to 70℃, stirring speed: 300 rpm, stirring time: 2 min; then homogenize, homogenization speed: 4000 rpm, homogenization time: 2 min, to obtain mixture B; S3: add mixture B to mixture A, stir, stirring speed: 300 rpm, stirring time: 5 min; then homogenize, homogenization speed: 4000 rpm, homogenization time: 5 min, to obtain mixture; S4: after the mixture is cooled to 40℃, mix the remaining deionized water with the mixture, stir, stirring speed: 300 rpm, stirring time: 2 min; then homogenize, homogenization speed: 2000 rpm, homogenization time: 2 min, continue to cool to 25℃, to obtain a blank emulsion, fill, sterilize, seal, and store at room temperature.

[0053] Test 1: Effect of the composition on elastase activity test The effect of the test sample on the activity of skin elastase is evaluated by an in vitro elastase activity inhibition test, and the change in enzyme activity is determined using a specific fluorescent substrate, thereby evaluating the anti-wrinkle firming effect.

[0054] The test samples include firming anti-aging compositions 1-3 and firming anti-aging compositions 1-10, and the specific grouping is as follows: Test sample group: 50 μL of the test sample solution prepared with the buffer in the kit at a concentration of 3 wt% is added to a black 96-well bottom transmittance plate, then 100 μL of 0.2 U / mL elastase working solution is added, and the reaction is shaken at room temperature for 15 min, then 50 μL of 100 μg / mL elastase fluorescent coupling compound is added to each well, and the fluorescence intensity at an excitation wavelength of 485 nm and an emission wavelength of 530 nm is measured after incubation at room temperature in the dark for 30 min, and is recorded as MFI 待测样品 ; Control group: the test sample solution is replaced with an equal amount of 50 μL buffer, and the rest remains unchanged, and the fluorescence intensity is measured and recorded as MFI 对照 ; Background group: the test sample solution is replaced with 50 μL buffer, and the elastase working solution is replaced with 100 μL buffer, and the rest remains unchanged, and the fluorescence intensity is measured and recorded as MFI 背景 ; The inhibition ability of different test samples on elastase activity is calculated according to the following formula: Elastase activity inhibition rate / % = [1- (MFI 待测样品 -MFI 背景 ) / (MFI 对照 -MFI 背景 )] x 100% The experimental results are shown in Table 2.

[0055] Table 2 Effects of the composition on the inhibition of elastase activity Group Elastase activity inhibition rate (%) Anti-aging composition 1 85.42 Anti-aging composition 2 84.39 Anti-aging composition 3 82.01 Anti-aging composition 1 57.22 Anti-aging composition 2 68.73 Anti-aging composition 3 62.67 Anti-aging composition 4 53.84 Anti-aging composition 5 60.45 Anti-aging composition 6 62.53 Anti-aging composition 7 65.26 Anti-aging composition 8 71.61 Anti-aging composition 9 70.31 Anti-aging composition 10 74.64 According to the results shown in Table 2, it can be seen that the elastase activity inhibition rate of the comparative firming anti-aging composition 1 and the firming anti-aging compositions 1-4 shows that there is a significant synergistic effect among the E. punctatum bark extract, the E. grandiflorum flower extract, the Tremella fuciformis extract, and the composite polypeptide; the results of the comparative firming anti-aging composition 1 and the firming anti-aging compositions 5-7 show that compared with the use of the carnosine, the acetyl tetrapeptide-5, and the tripeptide-10 citrulline alone, the use of the composite polypeptide with the limited mass ratio according to the present application has a better inhibition rate on the elastase activity; the results of the comparative firming anti-aging composition 1 and the firming anti-aging composition 8 show that when the composite polypeptide with the limited mass ratio according to the present application is used together with the E. punctatum bark extract, the E. grandiflorum flower extract, and the Tremella fuciformis extract, the composition has a better inhibition effect on the elastase activity; the results of the comparative firming anti-aging compositions 1-3 and the firming anti-aging composition 9 show that when the E. punctatum bark extract, the E. grandiflorum flower extract, the Tremella fuciformis extract, and the composite protease with the mass ratio according to the present application are used together, the composition has a better inhibition effect on the elastase activity; the results of the comparative firming anti-aging composition 1 and the firming anti-aging composition 10 show that compared with the Tremella fuciformis extract, when the Tremella fuciformis extract is used together with the composite polypeptide, the E. punctatum bark extract, and the E. grandiflorum flower extract, the composition has a better inhibition effect on the elastase activity.

[0056] Test 2: Effect of the composition on the collagenase activity The firming anti-aging compositions 1-3 and 1-10 were prepared into a sample solution with a concentration of 3 wt% using purified water, and the sample solution was prepared immediately before use.

[0057] The concentration of the Tricine buffer solution was 50 mmol / L, and the pH was 7.5.

[0058] A collagenase solution with a concentration of 0.8 U / mL and a FALGPA (N-[3-(2-furyl) acryloyl]-leucine-glycine-proline-alanine) solution with a concentration of 2 mmol / L were prepared using the Tricine buffer solution.

[0059] The reaction solution without the sample: 40 μL of purified water was mixed with 100 μL of the Tricine buffer solution, and then 20 μL of the collagenase solution with a concentration of 0.8 U / mL was added, and after mixing, the mixture was incubated at 25°C for 15 min, and then 40 μL of the FALGPA solution with a concentration of 2 mmol / L was added, and after 15 min, the absorbance value was measured at 335 nm, and the measurement was repeated three times in parallel, and the average value was taken.

[0060] Reaction solution without enzyme: 40 μL of sample solution was mixed with 100 μL of Tricine buffer, then 20 μL of Tricine buffer was added, mixed and incubated at 25°C for 15 min, then 40 μL of 2 mmol / L FALGPA solution was added, and the absorbance value was measured at 335 nm after 15 min. The determination was repeated three times, and the average value was taken.

[0061] Reaction solution without sample and enzyme: 40 μL of purified water was mixed with 100 μL of Tricine buffer, then 20 μL of Tricine buffer was added, mixed and incubated at 25°C for 15 min, then 40 μL of 2 mmol / L FALGPA solution was added, and the absorbance value was measured at 335 nm after 15 min. The determination was repeated three times, and the average value was taken.

[0062] Reaction solution with sample and enzyme: 40 μL of sample solution was mixed with 100 μL of Tricine buffer, then 20 μL of 0.8 U / mL collagenase solution was added, mixed and incubated at 25°C for 15 min, then 40 μL of 2 mmol / L FALGPA solution was added, and the absorbance value was measured at 335 nm after 15 min. The determination was repeated three times, and the average value was taken. The inhibition rate of the sample on collagenase was calculated according to the formula.

[0063] Inhibition rate / % = [1- (C-D) / (A-B)] x 100% In the formula: A is the absorbance of the reaction solution without sample; B is the absorbance of the reaction solution without sample and enzyme; C is the absorbance of the reaction solution with sample and enzyme; and D is the absorbance of the reaction solution without enzyme.

[0064] The experimental results are shown in Table 3.

[0065] Table 3 Effect of composition on collagenase activity Group Collagenase activity inhibition rate (%) Anti-aging composition 1 65.24 Anti-aging composition 2 62.64 Anti-aging composition 3 63.72 Anti-aging composition 1 36.31 Anti-aging composition 2 41.43 Anti-aging composition 3 45.62 Anti-aging composition 4 39.26 Anti-aging composition 5 49.21 Anti-aging composition 6 51.57 Anti-aging composition 7 50.91 Anti-aging composition 8 54.33 Anti-aging composition 9 52.29 Anti-aging composition 10 55.93 As shown in the results of Table 3, it can be seen from the comparison of the collagenase activity inhibition rate of firming and anti-aging composition 1 and firming and anti-aging compositions ①-④ that there is a significant synergistic effect among the E. japonica bark extract, E. grandiflorum flower extract, Tremella fuciformis extract, and the composite polypeptide in the composition; it can be seen from the comparison of the firming and anti-aging composition 1 and firming and anti-aging compositions ⑤-⑦ that the inhibition rate of the composite polypeptide with the limited mass ratio of the present application on the collagenase activity is higher than that of the use of the carnosine, acetyl tetrapeptide-5, and tripeptide-10 citrulline alone; it can be seen from the comparison of the firming and anti-aging composition 1 and firming and anti-aging composition ⑧ that the inhibition rate of the composition on the collagenase activity is higher when the composite polypeptide with the limited mass ratio of the present application is used together with the E. japonica bark extract, E. grandiflorum flower extract, and Tremella fuciformis extract; it can be seen from the comparison of the firming and anti-aging compositions 1-3 and firming and anti-aging composition ⑨ that the inhibition rate of the composition on the collagenase activity is better when the E. japonica bark extract, E. grandiflorum flower extract, Tremella fuciformis extract, and composite protease with the mass ratio of the present application are used together; it can be seen from the comparison of the firming and anti-aging composition 1 and firming and anti-aging composition ⑩ that the inhibition effect of the composition on the collagenase activity is better when the Tremella fuciformis extract is used together with the composite polypeptide, E. japonica bark extract, and E. grandiflorum flower extract than when the Tremella fuciformis extract is used alone.

[0066] Test 3: Human efficacy test 3-1. Human skin patch test Tested materials: emulsions 1-5.

[0067] Selection of subjects: healthy volunteers aged 21-57 years old were selected, who voluntarily participated and signed the informed consent, could complete the test as required during the test period, and were randomly divided into groups, with 30 subjects in each group.

[0068] Experimental method: qualified patch equipment was selected, and a closed patch test method was used. 2 mL of the tested material was placed in the patch equipment and evenly coated, and then an external low-sensitization adhesive tape was attached to the flexor of the forearm of the subject. After 24 hours, the tested material was removed, and the skin reaction was observed and recorded at 0.5, 24, and 48 hours after removal, respectively.

[0069] Result evaluation: the skin adverse reaction grading was determined according to the skin adverse reaction grading of the human trial test in the “Cosmetic Safety Technology Specification” (2015 edition), and the grading standard is shown in Table 4. The results are shown in Table 5.

[0070] Table 4 Skin adverse reaction grading standard Classification Phenomenon 0 level Skin has no reaction 1 level Skin appears pinkish red spots 2 level Skin appears red spots, infiltration and papules 3 level Skin appears red spots, edema, papules and blisters 4 level Skin appears red spots, edema and bullae Table 5 Results of human skin patch test

[0071] 3-2. Skin patch test of sensitive skin Tested materials: Emulsion 1-5, Emulsion 1-10.

[0072] Subject selection: Healthy volunteers aged 21-44 years old were selected, who met the positive lactic acid stinging test, voluntarily participated and signed the informed consent, could complete the test as required during the test period, and were randomly divided into groups, with 10 subjects in each group.

[0073] Experimental method: Volunteers randomly divided their bilateral faces into test and control sides. After cleansing, the product was evenly applied to the facial skin on the test side according to the product usage method, and no product was used on the control side. After 28 days of trial, the volunteers fed back the local skin reaction, and if there was an adverse reaction, a dermatologist evaluated the adverse reaction.

[0074] Result evaluation: The skin adverse reaction grading was determined according to the skin adverse reaction grading of human trial test in the “Cosmetic Safety Technology Specification” (2015 edition), and the grading standard is shown in Table 4, and the results are shown in Table 6.

[0075] Table 6 Results of human trial test

[0076] From the results of the human skin patch test, during the test period, emulsion 1-5 was safe and no adverse reactions were observed in the subjects.

[0077] From the results of the sensitive skin test, during the test period, emulsion 1-5 and emulsion 1-10 had no adverse reactions in the subjects, indicating that emulsion 1-5 and emulsion 1-10 were safe, mild and non-irritating, and suitable for sensitive skin.

[0078] 3-3 Skin elasticity test Test samples: Emulsion 1-5, Emulsion 1-10, and blank emulsion.

[0079] Test method: Volunteers with wrinkles and healthy skin without damage were selected, with an average age of 30-45 years old, and were randomly divided into 16 groups, with 10 in each group, and were used with the above test samples. The subjects applied 2g of emulsion on their faces, twice a day, for 30 consecutive days.

[0080] During the experiment, the subjects could not apply any other cosmetics to the experimental site. The Cutometer MPA580 skin elasticity instrument was used to collect data on day 0 and day 30, and the test site was the cheek. The probe was measured 3 times in the same test area, and the average value was taken.

[0081] The test parameter is R2 (the ratio of the amount of skin rebound Ua without negative pressure to the maximum amount of stretching Uf with negative pressure), the closer R2 is to 1, the better the elasticity of the skin, and the R2 change rate = (R2 value on day 30-R2 value on day 0) / R2 value on week 0. The test results are shown in Table 7.

[0082] Table 7 Effect of the composition on skin elasticity Group R2 value on day 0 R2 value on day 30 R2 change rate (%) Emulsion 1 0.553 0.688 24.41 Emulsion 2 0.517 0.634 22.63 Emulsion 3 0.586 0.731 24.74 Emulsion 4 0.561 0.693 23.53 Emulsion 5 0.532 0.655 23.12 Emulsion 1 0.575 0.652 13.39 Emulsion 2 0.547 0.631 15.36 Emulsion 3 0.563 0.641 13.85 Emulsion 4 0.548 0.619 12.96 Emulsion 5 0.524 0.598 14.12 Emulsion 6 0.536 0.602 12.31 Emulsion 7 0.567 0.641 13.05 Emulsion 8 0.511 0.597 16.83 Emulsion 9 0.542 0.627 15.68 Emulsion 10 0.539 0.635 17.81 Emulsion 11 Blank emulsion 0.525 0.553 5.33 As shown in the results in Table 7, it can be seen from the results of comparative emulsions 1-3, emulsions 1-10 and the blank emulsion that the emulsion containing the firming and anti-aging composition of the application has the effect of improving skin elasticity; it can be seen from the results of comparative emulsions 1-3 that when the addition amount of the firming and anti-aging composition in the emulsion is in the range of 1wt%-3wt%, the emulsion has the effect of improving skin elasticity, and when the addition amount of the firming and anti-aging composition in the emulsion is 2wt%, the cost performance is the highest; it can be seen from the results of comparative emulsion 1 and emulsions 4-5 that the emulsion obtained by compounding each raw material according to the limited amount ratio of the application has the effect of improving skin elasticity; it can be seen from the results of comparative emulsion 1 and emulsions 1-4 and 9 that when the Euphorbia tirucalli bark extract, Jasminum mesnyi flower extract, Auricularia auricula extract and composite polypeptide in the firming and anti-aging composition are compounded according to the specific amount ratio, the emulsion has a better effect of improving skin elasticity; it can be seen from the results of comparative emulsion 1 and emulsions 5-7 that the composition and amount ratio of the composite polypeptide in the firming and anti-aging composition will affect the effect of the emulsion on improving skin elasticity, and the emulsion using the composite polypeptide in the application has the best effect of improving skin elasticity; it can be seen from the results of comparative emulsion 1 and emulsion 10 that the types of raw materials in the firming and anti-aging composition will affect the effect of the composition, thereby affecting the effect of the emulsion.

[0083] The embodiments described above are part of the embodiments of the present application, rather than all the embodiments. The detailed description of the embodiments of the present application is not intended to limit the scope of the claimed application, but only represents selected embodiments of the present application. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor are within the scope of protection of the present application.

Claims

1. A firming anti-aging composition comprising an extract of the bark of the tree Euclea natalensis, characterized in that, The firming anti-aging composition comprises the following mass parts of components: Plectranthus verticillatus bark extract: 1-3 parts; Forsythia suspensa flower extract: 0.5-0.7 parts; Ganoderma applanatum extract: 2-4 parts; Complex polypeptide: 0.3-0.5 parts: The complex polypeptide comprises at least one of carnosine, acetyl tetrapeptide-5, and tripeptide-10 citrulline.

2. The firming anti-aging composition of claim 1, wherein The complex polypeptide is composed of carnosine, acetyl tetrapeptide-5, and tripeptide-10 citrulline in a mass ratio of 1:0.4-0.6:0.2-0.

6.

3. Use of the firming anti-aging composition according to claim 1 or 2 in the preparation of a cosmetic product having firming and anti-aging effects.

4. Use according to claim 3, characterized in that, The dosage form of the cosmetic product having firming and anti-aging effects is any one of a cream, a lotion, and an essence.

5. An emulsion having a firming, anti-aging efficacy, characterized in that, The lotion comprises 1-3 wt% of the firming anti-aging composition according to claim 1 or 2.

6. The emulsion having firming and anti-aging efficacy according to claim 5, wherein, The lotion further comprises the following mass percentage components: Humectant: 1-5 wt%; Emollient: 1-5 wt%; Emulsifier: 0.5-3 wt%; Preservative: 0.2-0.5 wt%; Deionized water is added to 100 wt%.

7. The emulsion having firming and anti-aging efficacy according to claim 6, wherein, The humectant is one or more of methylpropanediol, 1,3-propanediol, and butylene glycol; and the preservative is one or more of phenoxyethanol, sodium benzoate, p-hydroxyacetophenone, and sorbic acid.

8. The emulsion having firming and anti-aging efficacy according to claim 6, wherein, The emulsifier is one or more of C14-22 alcohol, C12-20 alkyl glucoside, and polyglyceryl-3 methyl glucose distearate.

9. The emulsion having firming and anti-aging efficacy according to claim 6, wherein, The emollient is one or more of squalane, white pool seed oil, shea butter, European hazel seed oil, and jojoba oil.

10. A method of preparing an emulsion having a firming, anti-aging efficacy according to any one of claims 6 to 9, characterized in that, The method comprises the following steps: S1: Mix the humectant and the preservative with 1 / 2 deionized water, heat to 60-80°C, stir at a speed of 100-300 rpm for 1-3 min, then homogenize at a speed of 3000-5000 rpm for 1-3 min to obtain a mixed solution A; S2: Mix the emollient and the emulsifier, heat to 60-80°C, stir at a speed of 200-400 rpm for 1-3 min, then homogenize at a speed of 3000-5000 rpm for 1-3 min to obtain a mixed solution B; S3: Add the mixed solution B to the mixed solution A, stir at a speed of 200-400 rpm for 3-5 min, then homogenize at a speed of 3000-5000 rpm for 3-5 min to obtain a mixed solution; S4: Mix the firming anti-aging composition with the remaining deionized water, heat to 30-40°C, stir at a speed of 100-300 rpm for 1-3 min to obtain a mixed solution C; S5: the temperature of the mixed solution is lowered to 30-50℃, the mixed solution C is mixed with the mixed solution, stirring, stirring speed: 100-300 rpm, stirring time: 1-3 min; then homogenization, homogenization speed: 1000-3000 rpm, homogenization time: 1-3 min, continue to lower the temperature to 20-25℃, to obtain the emulsion with the effect of firming and anti-aging, filling, sterilization, sealing, room temperature preservation.

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