A polypeptide anti-wrinkle composition and its application

Abstract

This invention relates to a polypeptide anti-wrinkle composition and its application. The components of the polypeptide anti-wrinkle composition include carnosine, acetyl hexapeptide-8, palmitoyl pentapeptide-4, and dipalmitoyl hydroxyproline. In this invention, the four components of carnosine, acetyl hexapeptide-8, palmitoyl pentapeptide-4, and dipalmitoyl hydroxyproline are rationally combined to have a synergistic effect. This not only improves the integrity of the skin barrier but also has a good ability to scavenge free radicals and promote collagen synthesis, restoring vitality and thus giving the composition a triple effect of repair, moisturizing, and anti-wrinkle.

Description

Technical Field

[0001] This invention belongs to the field of cosmetic technology, specifically relating to a polypeptide anti-wrinkle composition and its application. Background Technology

[0002] Skin aging is a physiological or pathological phenomenon characterized by degenerative changes in skin morphology and function caused by the combined effects of multiple factors. In aging skin, elasticity gradually decreases, and the skin becomes increasingly thinner, more fragile, and looser. Simultaneously, the skin barrier function weakens during the aging process, leading to dryness, flaking, and increased wrinkles. Therefore, to enhance the skin's own antioxidant capacity, reduce wrinkle formation, and delay skin aging, anti-wrinkle and anti-aging skincare products are gaining popularity.

[0003] CN104856928A discloses an anti-wrinkle and anti-aging cosmetic and its preparation method. The cosmetic contains an emulsifier, a moisturizer, a skin-nourishing agent, an active ingredient, and water. This skincare composition has a synergistic effect in improving skin elasticity, increasing skin elasticity, reducing fine lines and wrinkles on the face, and alleviating sagging, looseness, and wrinkles. It also features a lightweight texture and easy absorption.

[0004] CN109363990A discloses an anti-wrinkle and anti-aging skincare composition, its preparation method, and its application. This anti-wrinkle and anti-aging skincare composition includes: baobab fruit extract, adenosine, chamomile extract, hyaluronic acid solution, a polypeptide complex, and deionized water as the balance. The anti-wrinkle and anti-aging skincare composition provided by this invention is characterized by significant efficacy, gentle properties, non-irritation, and no side effects; the various ingredients have a synergistic effect, and when applied to a serum, it is suitable for all skin types, showing significant advantages in improving skin elasticity, reducing wrinkles, and increasing skin firmness.

[0005] However, the aforementioned anti-wrinkle and anti-aging cosmetics only offer preventative effects and are unlikely to achieve rapid and long-lasting improvements in the skin's anti-aging capabilities. Therefore, there is an urgent need to provide a more durable and effective anti-wrinkle and anti-aging product to meet people's needs. Summary of the Invention

[0006] In view of the shortcomings of the prior art, the present invention aims to provide a polypeptide anti-wrinkle composition and its application. The polypeptide anti-wrinkle composition can not only promote cell renewal and proliferation, but also stimulate the production of collagen and dermal matrix, and inhibit the production of metalloproteinases, thereby achieving the effect of improving wrinkles and delaying aging.

[0007] To achieve this objective, the present invention adopts the following technical solution:

[0008] In a first aspect, the present invention provides a polypeptide anti-wrinkle composition (the polypeptide anti-wrinkle composition is abbreviated as Caparo), wherein the components of the polypeptide anti-wrinkle composition (Caparo) include carnosine, acetyl hexapeptide-8, palmitoyl pentapeptide-4 and dipalmitoyl hydroxyproline.

[0009] In the peptide anti-wrinkle composition (Caparo) of the present invention, carnosine has the ability to chelate metal ions and can inhibit the lipid peroxidation catalyzed by copper ions by forming a complex with them. Carnosine also has the ability to capture hydroxyl radicals, elemental oxygen, and peroxide radicals, thus exhibiting good antioxidant effects. Acetyl hexapeptide-8 can inhibit the release of the neurotransmitter acetylcholine, weaken muscle contraction, thereby reducing wrinkles, and can also increase elastin activity, improve skin laxity, and make the skin smooth and firm. Palmitoyl pentapeptide-4 can stimulate the synthesis of type I and III collagen and fibronectin in fibroblasts, and can also stimulate the synthesis of soluble type IV collagen and mucopolysaccharides, thereby increasing skin elasticity and reducing wrinkles. Dipalmitoyl hydroxyproline plays a key role in collagen synthesis and stabilization, and can inhibit superoxide anions, awaken and repair the skin, thereby making the skin smooth and revitalized. The four components, when combined, have a synergistic effect, giving the composition a triple effect of repair, moisturizing, and anti-wrinkle.

[0010] Preferably, the components of the polypeptide anti-wrinkle composition include, by weight, 0.1-5 parts carnosine, 1-10 parts acetyl hexapeptide-8, 1-10 parts palmitoyl pentapeptide-4, and 0.1-3 parts dipalmitoyl hydroxyproline.

[0011] The amount of carnosine added in the polypeptide anti-wrinkle composition of the present invention can be 0.2 parts, 0.5 parts, 1 part, 1.5 parts, 2 parts, 2.5 parts, 3 parts, 3.5 parts, 4 parts or 4.5 parts, etc.;

[0012] The amount of acetyl hexapeptide-8 added can be 2 parts, 3 parts, 4 parts, 5 parts, 6 parts, 7 parts, 8 parts, or 9 parts, etc.

[0013] The amount of palmitoyl pentapeptide-4 added can be 2 parts, 3 parts, 4 parts, 5 parts, 6 parts, 7 parts, 8 parts, or 9 parts, etc.

[0014] The amount of dipalmitoyl hydroxyproline added can be 0.2 parts, 0.5 parts, 1 part, 1.5 parts, 2 parts, 2.5 parts, or 3 parts, etc.

[0015] In a second aspect, the present invention provides the application of the polypeptide anti-wrinkle composition as described in the first aspect in cosmetics.

[0016] Preferably, the cosmetic includes serum, lotion, emulsion, face cream, facial cleanser, or face mask.

[0017] Preferably, the polypeptide anti-wrinkle composition has a mass percentage of 1-20% in the cosmetic, for example, 2%, 5%, 8%, 10%, 12%, 15%, or 18%.

[0018] Thirdly, the present invention provides a serum comprising the polypeptide anti-wrinkle composition as described in the first aspect and deionized water.

[0019] Preferably, the essence further includes any one or a combination of at least two of the following: moisturizer, thickener, chelating agent, or emulsifier;

[0020] Preferably, the essence comprises, by weight, 1-20 parts of the polypeptide anti-wrinkle composition as described in the first aspect, 1-25 parts of moisturizer, 0.1-1 parts of chelating agent, 0.01-0.1 parts of thickener, 0.1-3 parts of emulsifier, and deionized water.

[0021] The amount of the polypeptide anti-wrinkle composition added to the essence of the present invention is 3 parts, 5 parts, 7 parts, 10 parts, 13 parts, 15 parts, 17 parts or 19 parts, etc.

[0022] The amount of the humectant added can be 3 parts, 5 parts, 7 parts, 10 parts, 13 parts, 15 parts, 17 parts, 20 parts, 22 parts, or 24 parts, etc.

[0023] The amount of the chelating agent added can be 0.2 parts, 0.3 parts, 0.4 parts, 0.5 parts, 0.6 parts, 0.7 parts, 0.8 parts, or 0.9 parts, etc.;

[0024] The amount of the thickener added can be 0.02 parts, 0.03 parts, 0.04 parts, 0.05 parts, 0.06 parts, 0.07 parts, 0.08 parts, or 0.09 parts, etc.

[0025] The amount of emulsifier added can be 0.2 parts, 0.5 parts, 1 part, 1.5 parts, 2 parts, 2.5 parts, or 3 parts, etc.

[0026] Preferably, the moisturizer includes any one or a combination of at least two of glycerin, butylene glycol, 1,2-hexanediol or 1,2-pentanediol.

[0027] Preferably, the chelating agent comprises sodium EDTA-2.

[0028] Preferably, the thickener comprises any one or a combination of at least two of hydroxyethyl acrylate / sodium acryloyldimethyl taurate copolymer, carbomer, or xanthan gum.

[0029] Preferably, the emulsifier comprises a combination of hydrogenated lecithin and PEG-40 hydrogenated castor oil.

[0030] In this invention, hydrogenated lecithin and PEG-40 hydrogenated castor oil are used in combination, which not only improves the emulsifying performance of the emulsifier, but also ensures the activity of the active ingredients in the serum, reduces the stickiness of use, balances the impact of the environment on the skin feel, and achieves a soothing and irritation-reducing effect.

[0031] Preferably, the mass ratio of hydrogenated lecithin to PEG-40 hydrogenated castor oil is 1:(0.5-3), for example, it can be 1:0.7, 1:1, 1:1.2, 1:1.5, 1:1.8, 1:2, 1:2.2, 1:2.5 or 1:2.8, etc.

[0032] Preferably, the serum further includes a skin conditioning agent.

[0033] Preferably, the skin conditioning agent has a mass percentage of 0.1-5% in the total amount of the serum, for example, it can be 0.2%, 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4% or 4.5%.

[0034] Preferably, the skin conditioning agent comprises carrageenan extract and / or ginger flower root extract.

[0035] Preferably, the skin conditioning agent is a combination of Carrageenan extract and ginger flower root extract.

[0036] In this invention, the skin conditioning agent, through the combination of Carrageenan extract and ginger flower root extract, can further enhance the skin's antioxidant capacity. At the same time, it synergistically enhances the anti-wrinkle composition, thereby increasing collagen synthesis, enhancing cell proliferation rate, and improving the integrity of the skin barrier. Furthermore, the combination of the two can reduce the secretion of TNF-α induced by ultraviolet radiation and reduce inflammatory response, thus making the essence more effective in removing wrinkles and anti-aging.

[0037] Preferably, the mass ratio of the *Chlorophytum comosum* extract to the ginger root extract is 1:(1-5), for example, it can be 1:1.5, 1:2, 1:2.5, 1:3, 1:3.5, 1:4 or 1:4.5, etc.

[0038] Preferably, the essence further includes any one or a combination of at least two of the following: a pH adjuster, a preservative, or a fragrance.

[0039] Fourthly, the present invention provides a method for preparing the serum as described in the third aspect, the method comprising: dissolving the polypeptide anti-wrinkle composition described in the first aspect in deionized water, mixing and stirring to obtain the serum.

[0040] Preferably, the preparation method includes:

[0041] (1) Add the polypeptide anti-wrinkle composition, moisturizer and thickener as described in the first aspect to deionized water and stir until completely dissolved to obtain an aqueous phase liquid;

[0042] (2) Add emulsifier, chelating agent and skin conditioning agent to the aqueous phase liquid obtained in step (1), mix and homogenize to obtain the essence liquid;

[0043] Preferably, the stirring temperature in step (1) is 40-50℃, for example, it can be 41℃, 42℃, 43℃, 44℃, 45℃, 46℃, 47℃, 48℃ or 49℃, etc.

[0044] Preferably, the temperature during the mixing and homogenization in step (2) is 60-70℃ (e.g., 61℃, 62℃, 63℃, 64℃, 65℃, 66℃, 67℃, 68℃ or 69℃, etc.), and the time is 0.5-1h (e.g., 0.5h, 0.6h, 0.7h, 0.8h, 0.9h or 1h, etc.).

[0045] Preferably, after homogenization in step (2), cooling is required, and the cooling temperature is 25-35℃, for example, 26℃, 27℃, 28℃, 29℃, 30℃, 31℃, 32℃, 33℃ or 34℃.

[0046] Preferably, step (2) further includes adding any one or a combination of at least two of a pH adjuster, a preservative, or a fragrance.

[0047] All the specific point values ​​within the range of the above values ​​can be selected, and will not be elaborated on here.

[0048] Compared with the prior art, the present invention has the following beneficial effects:

[0049] 1. In the peptide anti-wrinkle composition (Caparo) of the present invention, the four components of carnosine, acetyl hexapeptide-8, palmitoyl pentapeptide-4 and dipalmitoyl hydroxyproline are rationally combined to have a synergistic effect. It can not only improve the integrity of the skin barrier, but also has a good ability to scavenge free radicals and promote collagen synthesis, restore vitality, and thus give the composition a triple effect of repair, moisturizing and anti-wrinkle.

[0050] 2. The essence of the present invention, through the combination of the polypeptide anti-wrinkle composition and other components, can not only increase the nutrition of the skin and deeply moisturize the skin, but also has a good effect on firming the skin and reducing skin wrinkles.

[0051] 3. The essence preparation method provided by this invention is simple and easy to operate, easy to industrialize and mass-produce, and has excellent practicality. Detailed Implementation

[0052] To further illustrate the technical means and effects of the present invention, the following describes the technical solution of the present invention in conjunction with preferred embodiments of the present invention. However, the present invention is not limited to the scope of the embodiments.

[0053] The weight percentages of carnosine, acetyl hexapeptide-8, palmitoyl pentapeptide-4 and dipalmitoyl hydroxyproline mentioned below are based on the actual content of the active ingredients in commercially available raw materials. Commercially available raw materials may also selectively contain any one or a combination of at least two of the following: solvents, fillers, diluents, stabilizers, pH adjusters, antibacterial agents, antioxidants, or impurities within permissible limits.

[0054] The following are some of the material and grade information involved in the embodiments and comparative examples:

[0055]

[0056]

[0057] All other raw materials can be used as long as they are purchased from authorized distributors.

[0058] Example 1

[0059] This embodiment provides a polypeptide anti-wrinkle composition, which is prepared by the following method: 3 parts carnosine, 5 parts acetyl hexapeptide-8, 6 parts palmitoyl pentapeptide-4 and 2 parts dipalmitoyl hydroxyproline are mixed and homogenized to obtain the polypeptide anti-wrinkle composition.

[0060] Example 2

[0061] This embodiment provides a polypeptide anti-wrinkle composition, which is prepared by the following method: mixing 1 part carnosine, 8 parts acetyl hexapeptide-8, 3 parts palmitoyl pentapeptide-4 and 3 parts dipalmitoyl hydroxyproline to obtain the polypeptide anti-wrinkle composition.

[0062] Example 3

[0063] This embodiment provides a polypeptide anti-wrinkle composition, which is prepared by the following method: 5 parts of carnosine, 3 parts of acetyl hexapeptide-8, 8 parts of palmitoyl pentapeptide-4 and 1 part of dipalmitoyl hydroxyproline are mixed to obtain the polypeptide anti-wrinkle composition.

[0064] Example 4

[0065] This embodiment provides a polypeptide anti-wrinkle composition, which is prepared by the following method: 6 parts carnosine, 0.5 parts acetyl hexapeptide-8, 12 parts palmitoyl pentapeptide-4 and 0.1 parts dipalmitoyl hydroxyproline are mixed to obtain the polypeptide anti-wrinkle composition.

[0066] Example 5

[0067] This embodiment provides a polypeptide anti-wrinkle composition, which is prepared by the following method: 0.1 parts carnosine, 12 parts acetyl hexapeptide-8, 0.5 parts palmitoyl pentapeptide-4 and 5 parts dipalmitoyl hydroxyproline are mixed to obtain the polypeptide anti-wrinkle composition.

[0068] Comparative Example 1

[0069] This embodiment provides a polypeptide anti-wrinkle composition. The only difference between the polypeptide anti-wrinkle composition and that of Example 1 is that the composition does not include carnosine, and the reduced amount of carnosine is allocated to acetyl hexapeptide-8, palmitoyl pentapeptide-4 and dipalmitoyl hydroxyproline according to the ratio. The other steps are the same as those of Example 1.

[0070] Comparative Example 2

[0071] This embodiment provides a polypeptide anti-wrinkle composition. The only difference between the polypeptide anti-wrinkle composition and that of Example 1 is that the composition does not include acetyl hexapeptide-8, and the reduced amount of acetyl hexapeptide-8 is allocated to carnosine, palmitoyl pentapeptide-4 and dipalmitoyl hydroxyproline according to the ratio. The other steps are the same as those of Example 1.

[0072] Comparative Example 3

[0073] This embodiment provides a polypeptide anti-wrinkle composition. The only difference between the polypeptide anti-wrinkle composition and that of Example 1 is that the composition does not include palmitoyl pentapeptide-4, and the reduced amount of palmitoyl pentapeptide-4 is allocated to carnosine, acetyl hexapeptide-8 and dipalmitoyl hydroxyproline according to the ratio. The other steps are the same as those of Example 1.

[0074] Comparative Example 4

[0075] This embodiment provides a polypeptide anti-wrinkle composition. The only difference between the polypeptide anti-wrinkle composition and that of Example 1 is that it does not include dipalmitoyl hydroxyproline in the components, and the reduced amount of dipalmitoyl hydroxyproline is allocated to carnosine, acetyl hexapeptide-8 and palmitoyl pentapeptide-4 according to the ratio. The other steps are the same as those of Example 1.

[0076] Application Example 1

[0077] This application example provides a serum with the following formula:

[0078]

[0079] Its preparation method is as follows:

[0080] (1) Add the anti-aging composition, moisturizer and thickener obtained in Example 1 to deionized water, stir until completely dissolved, and let stand for 4 hours to obtain an aqueous phase liquid;

[0081] (2) Add emulsifier, antioxidant and skin conditioning agent to the aqueous phase obtained in step (1), mix and homogenize at 80°C for 20 min, and cool to 30°C to obtain the essence.

[0082] Application Example 2-5

[0083] This application example provides four serums, the only difference between which is that the peptide anti-wrinkle composition of Example 1 is replaced with the peptide anti-wrinkle compositions of Examples 2-5 in the same amount, while all other conditions remain unchanged, and the preparation method is the same as in Application Example 1.

[0084] Application Example 6

[0085] This application example provides a serum whose formula differs from Application Example 1 only in that it does not contain a skin conditioning agent; the reduced amount is made up by water, while all other conditions remain unchanged. Its preparation method is the same as in Application Example 1.

[0086] Application Example 7

[0087] This application example provides a serum whose formula differs from Application Example 1 only in that the skin conditioning agent is ginger flower root extract, and the amount of skin conditioning agent added remains unchanged, while all other conditions remain the same. Its preparation method is the same as in Application Example 1.

[0088] Application Example 8

[0089] This application example provides a serum whose formula differs from Application Example 1 only in that the skin conditioning agent is *Chlorella vulgaris* extract, and the amount of skin conditioning agent added remains unchanged, while all other conditions remain the same. Its preparation method is the same as in Application Example 1.

[0090] Application Example 9

[0091] This application example provides an essence, the only difference in formulation from Application Example 1 being that the emulsifier is hydrogenated lecithin, and the amount of emulsifier added remains unchanged, while all other conditions remain the same. Its preparation method is the same as in Application Example 1.

[0092] Application Example 10

[0093] This application example provides an essence whose formulation differs from Application Example 1 only in that the emulsifier is PEG-40 hydrogenated castor oil, and the amount of emulsifier added remains unchanged, while all other conditions remain the same. Its preparation method is the same as in Application Example 1.

[0094] Compare and contrast examples 1-4

[0095] This application example provides four types of serums, the only difference between which are the formulations and those of Application Example 1. The peptide anti-wrinkle composition of Example 1 is replaced with the peptide anti-wrinkle compositions of Comparative Examples 1-3 in the same amount, while all other conditions remain unchanged. The preparation method is the same as in Application Example 1.

[0096] Comparative Application Example 5

[0097] This application example provides a serum whose formula differs from Application Example 1 only in that it does not contain the peptide anti-wrinkle composition; the reduction is made up by water, while all other conditions remain unchanged. Its preparation method is the same as in Application Example 1.

[0098] Test Example 1: Antioxidant Efficacy Study – Free Radical Scavenging Experiment

[0099] Free radical scavenging tests were performed on the serums provided in corresponding use cases 1-10 and application comparison examples 1-5, using the following method:

[0100] (1) Determination of superoxide anion free radical scavenging ability

[0101] Take 4.5 mL of 0.05 mol / L Tris-HCl buffer solution with pH 8.2, preheat it in a 25°C water bath for 20 min, then add 1 mL of the sample to be tested, followed by 0.4 mL of 25 mmol / L pyrogallol solution. Mix well and react in a 25°C water bath for 5 min. Finally, add 1.0 mL of 8 mol / L HCl to terminate the reaction.

[0102] Using Tris-HCl buffer as a reference, absorbance was measured at 299 nm. A blank control was prepared by replacing the sample with 1 mL of distilled water, and the clearance rate was calculated. The clearance rate formula is (D)(%) = [1 - (A4 / A3)] × 100%; where A3 is the absorbance value of the blank control; and A4 is the absorbance value of the sample.

[0103] (2) Determination of scavenging ability against hydroxyl radicals

[0104] Add 3 mL of 2 mmol / L FeSO4 and 3 mL of 1 mmol / L H2O2 to a 25 mL colorimetric tube, shake well, then add 3 mL of 6 mmol / L salicylic acid, shake well, heat in a 37°C water bath for 15 min, and then measure the absorbance A. o Then add 100 mg·L⁻¹ respectively. -1 Take the sample to be tested, shake well, continue heating in a water bath for 15 minutes, and then measure its absorbance A. x Following the above method, take another colorimetric tube, and repeat the process of adding each of the above liquids in sequence, without adding salicylic acid. Shake well, heat in a water bath for 15 minutes, and measure value A. xoThe formula for calculating the hydroxyl radical scavenging rate is: Hydroxyl radical scavenging rate (%) = A0 - (A0 - A0) / ( ... x -A x0 ) / A0×100%; where A0 is the absorbance value of the blank control system; A x The absorbance value of the sample system to be tested; A x0 The absorbance value is the background value of the solution without the colorimetric reagent H2O2.

[0105] The specific test results are shown in Table 1:

[0106] Table 1

[0107]

[0108]

[0109] As shown in Table 1, the serums provided in Application Examples 1-3 of this invention exhibit a scavenging rate of over 70% for superoxide anion free radicals and an inhibition rate of over 69% for hydroxyl free radicals, demonstrating excellent free radical scavenging and antioxidant effects. Application Examples 1 and 9-10 show that the combination of hydrogenated lecithin and PEG-40 hydrogenated castor oil used in this invention as an emulsifier provides better emulsification performance and better preserves the activity of the active ingredients. Application Examples 1 and Comparative Application Examples 1-4 show that the absence of any one of carnosine, acetyl hexapeptide-8, palmitoyl pentapeptide-4, and dipalmitoyl hydroxyproline affects the product's antioxidant effect. Comparative Application Example 4 shows that when the product does not contain a polypeptide anti-wrinkle composition, the final antioxidant performance is significantly poor.

[0110] Test Example 2: Wrinkle Reduction and Skin Firming Test

[0111] The skin-firming effect of the serums prepared in corresponding use cases 1-10 and comparative application examples 1-5 was evaluated:

[0112] 150 participants aged 20-45 were randomly selected and divided into 15 groups of 10 each. The 15 groups of participants used the serums prepared in Application Examples 1-10 and Control Application Examples 1-5 (twice daily, morning and evening) for 30 days. During the trial, participants were not allowed to use other products that maintain skin elasticity or cosmetics that could affect the results, and their original dietary habits were not changed.

[0113] Skin condition was tested on day 0 and day 30 using a Cutometer MPA580 skin elastometer. The test site was the cheekbone area. Measurements were taken three times on the same area, and the average value was recorded. The test parameter was R² (the ratio of skin rebound Ua without negative pressure to the maximum stretch Uf under negative pressure). The closer R² is to 1, the better the skin elasticity. The average change in R² = R² value on day 30 - R² value on day 0, and the average rate of change in R² = average change in R² / R² value on day 0. The specific skin elasticity test results are shown in Table 2.

[0114] Table 2

[0115]

[0116]

[0117] The results showed that the essence involved in this invention has a good skin-tightening effect. Among them, the four components, carnosine, acetyl hexapeptide-8, palmitoyl pentapeptide-4 and dipalmitoyl hydroxyproline, promote each other in this effect and have an unexpected synergistic effect. In addition, the effect is better when the polypeptide anti-wrinkle composition is used in combination with skin conditioning agents.

[0118] Test Example 3: VISIA Skin Analysis and Evaluation Experiment

[0119] The VISIA skin image analyzer employs multispectral imaging technology, using a 12-megapixel camera to capture images from three angles: front, left, and right. It images three times under three light sources: standard white light, cross-sectional polarized light, and ultraviolet light. Therefore, the VISIA facial image analysis system can systematically analyze the pigmentation, texture, pores, wrinkles, ultraviolet spots, stable pigmentation, red areas, and purplish-red areas of the subject's facial skin, providing precise and quantitative analytical data. By recording the pigmentation, wrinkles, and texture of the subject's face before and after the test, the absolute score represents the area and intensity of skin characteristic detection values, used to observe changes in skin properties. Therefore, the data obtained from the absolute score (the average of the test results from the front, left, and right sides) is selected as the analytical standard and data statistical basis. All data for each subject are entered into a computer, the measurement results are compared, and statistical analysis is performed to analyze the subject's skin characteristics and various indicator data. The testing instrument used in the experiment is the VISIA facial image analysis system (CANFIELD Technologies, USA).

[0120] The specific testing method is as follows:

[0121] Ninety women aged 30-40 with aging skin were randomly divided into nine groups of ten each. Each group used the serum prepared in Example 1, Examples 6-8, and the control examples 1-5, respectively. Participants were required to use the serum continuously, applying it twice daily (morning and evening) for 28 consecutive days. Follow-up visits were required on day 14 (D14) and day 28 (D28). The same participant's test was conducted by the same measurement personnel.

[0122] No products (cosmetics, topical medications, or oral health supplements) should be used on the test area for 15 days prior to the test. After thoroughly cleansing the face with a standardized facial cleanser, gently pat dry with a tissue. In a room with a temperature of 22℃±2℃ and a relative humidity of 40±2%, wait 15 minutes before using the VISIA facial image analyzer to photograph the subject's face. The analysis area should be kept as clear as possible, avoiding reflective areas and interference from hair. Photographs of the subject's face (including the front, left side, and right side) are taken. The absolute scores are statistically analyzed, and the detection data for facial texture and wrinkles are recorded. The test results are shown in Table 3.

[0123] Table 3

[0124]

[0125]

[0126] Based on the above experimental results, it can be seen that Application Example 1 has a significant effect on improving the wrinkles and texture of the subjects, indicating that the essence combination provided by the present invention can achieve a good effect in removing wrinkles and smoothing texture. However, the improvement effect of Application Example 6 is reduced, indicating that the skin conditioning agent and the peptide anti-wrinkle composition are used together for better results. As can be seen from Application Examples 7-8, the type of skin conditioning agent also affects the anti-wrinkle effect of the essence. The decrease in improvement effect in Application Examples 1-4 indicates that the four components, carnosine, acetyl hexapeptide-8, palmitoyl pentapeptide-4, and dipalmitoyl hydroxyproline, promote each other in improving wrinkles and texture and have a synergistic effect. As can be seen from Application Example 5, when the peptide anti-wrinkle composition of the present invention is lacking in the essence, it has a significant negative impact on the effect of the essence in improving wrinkles.

[0127] Test Example 4: Safety Evaluation Experiment

[0128] This test case involves a safety test of the serum combination prepared according to test cases 1-10. The test method is as follows:

[0129] The chorioallantoic membrane (CAM) is a respiratory membrane surrounding the chicken embryo. Because the surface of the CAM is richly vascularized, it can be considered a complete organism. This experiment utilizes the intact, clear, and transparent vascular system of the mid-stage chorioallantoic membrane in hatched chicken embryos. A certain amount of test substance was directly contacted with the chorioallantoic membrane, and after a specified exposure time, changes in chorioallantoic membrane toxicity indicators (such as hemorrhage, coagulation, and vascularization) were observed, scored, and the mathematical average was calculated to assess the irritant properties of the test substance.

[0130] (1) Chicken embryo preparation: Fertilized chicken embryos of breeds such as White Leghorn chicken, weighing 50g-60g, preferably SPF chicken embryos, are placed in an incubator for incubation. The temperature is controlled at 37.5℃±0.5℃, the relative humidity is 55%-70%, and the eggs are turned 3-6 times / h. On the 8th day of incubation, the eggs are checked with an egg candler. Unfertilized and inactive chicken embryos are discarded, and chicken embryos with well-developed blood vessels are selected. The location of the air cell is marked on the eggshell surface.

[0131] (2) Testing Procedure: At least 6 chicken embryos were used in each group, and negative and positive controls were required for each experiment. The experiment began on day 9. The air cell shell was carefully removed using dental tweezers. A few drops of physiological saline were applied to the surface of the shell membrane to fully moisten it. After pouring out the saline, the shell membrane was carefully removed with tweezers, ensuring that the exposed allantoic membrane remained intact without any damage. 0.3 ml / 0.3 g of the test substance was applied directly to the CAM. It was spread as widely as possible to ensure that the coverage area reached at least 50%. After 3 minutes of application, the CAM surface was gently rinsed with double-distilled water within 30 seconds. The liquid was poured out, and the degree of change of each toxic effect was immediately observed under a stereomicroscope. The toxic effects were scored based on capillary bleeding, coagulation, and vascularization (ES).

[0132] (3) Results observation:

[0133] Bleeding: Blood flows out from the blood vessels or capillaries of the CAM. Bleeding is scored as 0, 1, 2, or 3 points respectively, based on no bleeding, mild bleeding, moderate bleeding, and severe bleeding.

[0134] Coagulation: refers to the denaturation of proteins inside and outside blood vessels, manifested as thrombosis, swelling of the vessel wall, appearance of coagulation points inside and outside blood vessels, and milky opacity outside the blood vessels. It is scored as 0, 1, 2, and 3 points respectively based on no coagulation, mild coagulation, moderate coagulation, and severe coagulation.

[0135] Vascular ablation: refers to the ablation of blood vessels on the CAM membrane. A score of 0, 1, 2, and 3 is assigned based on whether there is no vascular ablation, mild vascular ablation, moderate vascular ablation, or severe vascular ablation, respectively.

[0136] (4) Result Calculation and Evaluation

[0137] Calculate the average ES score using the following formula, and classify the stimuli according to Table 4 below.

[0138]

[0139] Table 4. Results Evaluation

[0140] Finish line score Irritant Classification ES≤12 Non-irritating / mildly irritating 12<ES<16 moderate irritation ES≥16 Severe irritation

[0141] The results are shown in Table 5:

[0142] Table 5

[0143] result Application Example 1 0 Application Example 2 0 Application Example 3 0 Application Example 4 0 Application Example 5 0 Application Example 6 0 Application Example 7 0 Application Example 8 0 Application Example 9 0 Application Example 10 0

[0144] As shown in Table 5, the essence of the present invention does not irritate the skin and has no positive reaction, indicating that the product of the present invention is safer and gentler.

[0145] The applicant declares that the present invention illustrates a polypeptide anti-wrinkle composition and its application through the above embodiments, but the present invention is not limited to the above embodiments, that is, it does not mean that the present invention must rely on the above embodiments to be implemented. Those skilled in the art should understand that any improvements to the present invention, equivalent substitutions of the raw materials of the product of the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

[0146] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solution of the present invention, and these simple modifications all fall within the protection scope of the present invention.

[0147] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not describe the various possible combinations separately.

Claims

1. An essence, characterized in that, The serum is composed of a polypeptide anti-wrinkle composition, a moisturizer, a thickener, a chelating agent, an emulsifier, a skin conditioning agent, and deionized water; the polypeptide anti-wrinkle composition consists of 1-5 parts carnosine, 3-8 parts acetyl hexapeptide-8, 3-8 parts palmitoyl pentapeptide-4, and 1-3 parts dipalmitoyl hydroxyproline. The skin conditioning agent is a combination of Chondrus crispus extract and ginger flower root extract; The mass ratio of the *Chlorophytum comosum* extract to the ginger root extract is 1:(1-5). The emulsifier is a combination of hydrogenated lecithin and PEG-40 hydrogenated castor oil in a mass ratio of 1:(0.5-3).

2. The essence as described in claim 1, characterized in that, The peptide anti-wrinkle composition has a mass percentage of 1-20% in the serum.

3. The serum as described in claim 1, characterized in that, The essence is composed of 1-20 parts by weight of the aforementioned polypeptide anti-wrinkle composition, 1-25 parts by weight of moisturizer, 0.01-0.1 parts by weight of chelating agent, 0.1-1 parts by weight of thickener, 0.1-3 parts by weight of emulsifier, skin conditioning agent and deionized water.

4. The essence as described in claim 1, characterized in that, The moisturizer includes any one or a combination of at least two of glycerin, butylene glycol, 1,2-hexanediol, or 1,2-pentanediol.

5. The serum as described in claim 1, characterized in that, The chelating agent includes sodium EDTA-2.

6. The serum as described in claim 1, characterized in that, The thickener includes any one or a combination of at least two of the following: hydroxyethyl acrylate / sodium acryloyl dimethyl taurate copolymer, carbomer, or xanthan gum.

7. The serum as described in claim 1, characterized in that, The skin conditioning agent comprises 0.1-5% by mass of the total amount of serum.

8. The method for preparing the serum according to any one of claims 1-7, characterized in that, The preparation method includes: dissolving the polypeptide anti-wrinkle composition in deionized water, mixing and stirring to obtain the essence.

9. The method for preparing the essence as described in claim 8, characterized in that, The preparation method includes: (1) Add the peptide anti-wrinkle composition, moisturizer and thickener to deionized water and stir until completely dissolved to obtain an aqueous phase liquid; (2) Add emulsifier, chelating agent and skin conditioning agent to the aqueous phase obtained in step (1), mix and homogenize to obtain the essence.

10. The method for preparing the essence as described in claim 9, characterized in that, The stirring temperature in step (1) is 40-50℃.

11. The method for preparing the essence as described in claim 9, characterized in that, The mixing and homogenization process in step (2) is carried out at a temperature of 60-70℃ for 0.5-1h.

12. The method for preparing the essence as described in claim 9, characterized in that, After homogenization in step (2), cooling is required, specifically cooling to 25-35℃.

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