Instant anti-wrinkle polypeptide composition as well as preparation method and application thereof

Abstract

The invention discloses an instant anti-wrinkle polypeptide composition as well as a preparation method and application thereof, and belongs to the technical field of cosmetics. The polypeptide in the polypeptide composition comprises acetyl hexapeptide-8, acetyl octapeptide-3, dipeptide diaminobutyryl benzyl amide diacetate, arginine / lysine polypeptide and palmitoyl pentapeptide-4. The invention further discloses a preparation method of the polypeptide composition. The polypeptide composition disclosed by the invention can effectively inhibit release of acetylcholine, so that expression lines are quickly smoothed, and an instant anti-wrinkle effect is realized; meanwhile, the stability of the anti-wrinkle effect is maintained, wrinkles are prevented from rebounding, skin aging is delayed, the skin elasticity is cooperatively improved, and the skin is instantly smooth, elastic, moist and full.

Description

Technical Field

[0001] This invention relates to the field of cosmetic technology, specifically to an instant anti-wrinkle polypeptide composition, its preparation method, and its application. Background Technology

[0002] Skin aging is a complex physiological process, manifested as increased wrinkles, decreased elasticity, dryness, roughness, and dullness. Its causes include endogenous factors such as genetics and cell metabolism, and exogenous factors such as ultraviolet radiation and pollution. At the molecular level, it is mainly related to the reduced synthesis and accelerated degradation of extracellular matrix components such as collagen and elastin in the dermis, as well as the repetitive contractions of facial muscles due to over-excitation at the neuromuscular junction.

[0003] Among numerous anti-aging ingredients, peptides have become a research hotspot in the field of functional cosmetics due to their high efficacy and gentleness. Peptides are short chains formed by amino acids linked by peptide bonds, capable of targeting specific physiological sites in the skin. Their main anti-wrinkle mechanisms include: as signal peptides, stimulating fibroblasts, promoting the synthesis of collagen and elastin, and improving skin firmness and elasticity, such as palmitoyl pentapeptide-4; as neurotransmitter inhibitory peptides, intervening in the SNARE complex, inhibiting the release of acetylcholine, relaxing facial muscles, and thus reducing the formation of dynamic wrinkles, such as acetyl hexapeptide-8; and as carrier peptides, promoting the transport of trace elements (such as copper ions) and participating in the skin repair process, such as tripeptide-1 copper.

[0004] Currently, in the field of cosmetic technology applications, there are many application cases of skin application solutions for peptides and their compositions. For example, CN113116742A discloses a compound peptide anti-wrinkle combination containing the following components: 0.05-3% acetyl tetrapeptide-5, 0.05-2% palmitoyl tetrapeptide-7, 0.1-3% acetyl hexapeptide-8, 0.1-3% snake venom peptide, and 0.1-3% sodium hyaluronate. This patent focuses on enhancing skin elasticity and delaying skin aging through the physical mixing of peptide raw materials, but its effects are long-lasting, with only 28 days of moisturizing efficacy. CN110179687A discloses a multi-effect repair peptide composition containing multiple repair ingredients such as oligopeptide-1, carnosine, copper peptide, palmitoyl tetrapeptide-7, and nonapeptide-1. This patent focuses on moisturizing, anti-oxidation, spot fading, anti-wrinkle, and soothing sensitive irritation effects, but the accumulation of ingredients may lead to complex formulations, potential interactions between ingredients affecting stability, and higher costs. Furthermore, some products suffer from shortcomings such as limited effectiveness and high irritation, failing to meet consumers' actual needs. Achieving "instant anti-wrinkle" through scientific compounding techniques remains a significant challenge for the industry. Therefore, there is an urgent need to provide an anti-wrinkle and anti-aging product with a clear mechanism, well-defined targets, and immediate effects to meet people's needs. Summary of the Invention

[0005] The purpose of this invention is to provide an instant anti-wrinkle polypeptide composition, its preparation method and application, which effectively inhibits the release of acetylcholine, thereby quickly smoothing expression lines and achieving an instant anti-wrinkle effect; at the same time, it maintains the stability of the anti-wrinkle effect, prevents wrinkle rebound, delays skin aging, and synergistically improves skin elasticity, so that the skin can achieve a dual improvement of anti-wrinkle smoothness and elasticity.

[0006] The technical solution adopted by the present invention to achieve the above objectives is as follows: An instant anti-wrinkle polypeptide composition comprising a polypeptide and a preservative; the polypeptide comprises at least one selected from acetyl hexapeptide-8, acetyl octapeptide-3, dipeptide diaminobutyryl benzylamide diacetate, arginine / lysine polypeptide, and palmitoyl pentapeptide-4; the polypeptide composition exhibits an acetylcholine inhibition rate of 43.53-57.49% over a 2-hour treatment period.

[0007] This invention combines acetyl hexapeptide-8, acetyl octapeptide-3, dipeptide-diaminobutyryl benzylamide diacetate, arginine / lysine peptide, and palmitoyl pentapeptide-4 to form an immediate anti-wrinkle peptide composition. Acetyl hexapeptide-8 and acetyl octapeptide-3 act on the presynaptic membrane, competitively inhibiting SNAP-25 protein and mimicking its N-terminal structure, respectively, to interfere with the formation of the SNARE complex and effectively inhibit the release of the neurotransmitter acetylcholine. Dipeptide-diaminobutyryl benzylamide diacetate and arginine / lysine peptide act on the postsynaptic membrane, respectively antagonizing the ε subunit of the acetylcholine receptor (nmAChR) and inhibiting voltage-gated sodium ion channels, thereby blocking the transmission of muscle contraction signals and reducing fine lines. Palmitoyl pentapeptide-4, as a signal peptide, promotes the synthesis of extracellular matrix such as collagen to improve the skin's supporting structure. The polypeptide composition disclosed in this invention targets key points in the process of expression line formation, from reducing the release of the neurotransmitter acetylcholine that causes muscle contraction at the presynaptic membrane, to inhibiting nerve impulse transmission at the postsynaptic membrane, and finally promoting collagen production, thereby achieving an immediate anti-wrinkle effect.

[0008] Preferably, the polypeptide comprises the following components in parts by weight: 30-50 parts acetyl hexapeptide-8, 1-20 parts acetyl octapeptide-3, 30-50 parts dipeptide diaminobutyryl benzylamide diacetate, 1-20 parts arginine / lysine polypeptide and 0.1-10 parts palmitoyl pentapeptide-4.

[0009] Preferably, the preservative comprises 0.5-2 parts by weight of ethylhexylglycerin and 10-30 parts by weight of 1,2-hexanediol.

[0010] Preferably, the mass ratio of polypeptide to preservative is 1:50-200.

[0011] Preferably, the polypeptide composition includes a polypeptide, a functional additive, and a preservative, wherein the functional additive is 23-hydroxybetulic acid.

[0012] More preferably, the mass ratio of peptide to preservative is 1:50-200.

[0013] More preferably, the mass ratio of peptide to functional additive is 1:0.1-1.

[0014] Preferably, the functional additive comprises at least two of the following components in parts by weight: 2-5 parts of 23-hydroxybetulic acid, 0.6-1.4 parts of isoflavone acid, 3.5-4.6 parts of modified gallic acid ester, and 5-20 parts of ethanol.

[0015] More preferably, the functional additive comprises the following components in parts by weight: 2-5 parts of 23-hydroxybetulic acid and 5-20 parts of ethanol. The use of 23-hydroxybetulic acid in the peptide composition of this invention may effectively inhibit the release of the neurotransmitter acetylcholine by affecting the stability of the SNARE complex, thereby alleviating muscle contraction and effectively improving dynamic wrinkles; simultaneously, it promotes collagen synthesis, enhances skin barrier function, strengthens the skin's intrinsic support and elasticity, and achieves stable anti-wrinkle effects.

[0016] More preferably, the functional additive comprises the following components in parts by weight: 2-5 parts 23-hydroxybetulic acid, 0.6-1.4 parts usnic acid, and 5-20 parts ethanol. The use of usnic acid in the polypeptide composition of this invention may, through its antioxidant effect, scavenge excess free radicals, protect the existing collagen structure within cells from damage, thereby maintaining and improving skin elasticity and anti-wrinkle effects.

[0017] More preferably, the functional additive comprises the following components in parts by weight: 2-5 parts 23-hydroxybetulic acid, 0.6-1.4 parts usnic acid, 5-20 parts modified gallic acid ester, and 5-20 parts ethanol. The present invention uses modified gallic acid ester in peptide compositions, which may neutralize free radicals to resist oxidative stress damage to structural proteins and promote collagen synthesis to rebuild the skin's supporting structure; simultaneously improving the dynamic microenvironment of the skin, thereby comprehensively enhancing skin elasticity and anti-wrinkle effects.

[0018] More preferably, the method for preparing modified gallic acid esters includes, S1. Gallic acid and isobutyric anhydride are mixed and dissolved in N,N-dimethylformamide. 4-Dimethylaminopyridine and triethylamine are added and stirred for 1-5 hours. The reaction is terminated with dilute hydrochloric acid. The mixture is washed 2-5 times with deionized water and filtered to obtain triisobutyrylgalic acid.

[0019] S2. Triisobutyrylgalic acid and 1,3-dioxane-2-methanol were mixed and dissolved in N,N-dimethylformamide. 4-Dimethylaminopyridine and N,N'-dicyclohexylcarbodiimide were added and stirred for 3 h. The reaction was terminated with dilute hydrochloric acid. The mixture was extracted three times with ethyl acetate, and the organic phase was retained and dried over anhydrous magnesium sulfate. The mixture was purified by vacuum distillation and column chromatography to obtain modified gallic ester.

[0020] More preferably, the molar ratio of gallic acid to isobutyric anhydride in step S1 is 1:1-5.

[0021] More preferably, in step S1, the ratio of gallic acid to N,N-dimethylformamide is 1 mmol: 2-10 mL.

[0022] More preferably, the molar ratio of gallic acid and 4-dimethylaminopyridine in step S1 is 1:0.05-0.2.

[0023] More preferably, the molar ratio of gallic acid to triethylamine in step S1 is 1:3-6.

[0024] More preferably, the concentration of dilute hydrochloric acid in step S1 is 0.5-2 mol / L.

[0025] More preferably, in step S1, the volume ratio of N,N-dimethylformamide to dilute hydrochloric acid is 1:0.5-2.

[0026] More preferably, in step S1, the volume ratio of N,N-dimethylformamide to deionized water is 1:0.5-2.

[0027] More preferably, in step S2, the molar ratio of triisobutyrylgalic acid to 1,3-dioxane-2-methanol is 1:0.2-1.

[0028] More preferably, in step S2, the ratio of triisobutyrylgalic acid to N,N-dimethylformamide is 1 mmol: 2-10 mL.

[0029] More preferably, in step S2, the molar ratio of triisobutyrylgalic acid to 4-dimethylaminopyridine is 1:0.05-0.2.

[0030] More preferably, in step S2, the molar ratio of triisobutyrylgalic acid and N,N'-dicyclohexylcarbodiimide is 1:0.5-2.

[0031] More preferably, the concentration of dilute hydrochloric acid in step S2 is 0.5-2 mol / L.

[0032] More preferably, in step S2, the volume ratio of N,N-dimethylformamide to dilute hydrochloric acid is 1:0.5-2.

[0033] More preferably, in step S2, the volume ratio of N,N-dimethylformamide to ethyl acetate is 1:0.5-2.

[0034] The present invention also discloses the application of the above-mentioned instant anti-wrinkle polypeptide composition in the preparation of cosmetics.

[0035] Preferably, the cosmetic comprises an aqueous phase and an additive phase.

[0036] More preferably, the aqueous phase comprises the following components in parts by weight: 1-5 parts glycerol, 1-5 parts butanediol, 0.1-0.5 parts p-hydroxyacetophenone, 0.1-1 part 1,2-hexanediol, 1-3 parts 1,2-pentanediol and 0.01-0.05 parts disodium ethylenediaminetetraacetate, with the remainder being deionized water.

[0037] More preferably, the additive phase comprises 0.1-5 parts glycerol, 0.01-1 part 1,2-hexanediol, 0.001-0.01 parts PPG-5-cetyl ether-20, 0.001-0.01 parts ethylhexylglycerol and 0.05-20 parts by weight of the polypeptide composition, with the remainder being deionized water.

[0038] More preferably, the mass ratio of the aqueous phase to the additive phase is 1:0.01-0.4.

[0039] Preferably, the preparation method of the cosmetic is as follows: S1. Weigh each component according to the mass fractions of butanediol and p-hydroxyacetophenone in the aqueous phase. Mix glycerol, butanediol and p-hydroxyacetophenone, and heat to 55-75℃ while stirring. Stir until the mixture is clear and free of particles to obtain a clear, particle-free liquid for later use.

[0040] S2. Weigh each component according to the mass fractions of glycerol, 1,2-hexanediol, 1,2-pentanediol, disodium EDTA, and deionized water in the aqueous phase. Stir 1,2-hexanediol, 1,2-pentanediol, and disodium EDTA until there are no particles. Then, add 20-30% of the total volume of deionized water and stir until homogeneous. Add the remaining deionized water and stir until homogeneous to obtain a transparent liquid for later use.

[0041] S3. Add the transparent liquid obtained in step S2 to the transparent, particle-free liquid obtained in step S1, keep it at 55-75℃ for 10-30 minutes, then cool it to room temperature, add the additive phase and mix to obtain the cosmetic.

[0042] This invention combines five peptides—acetyl hexapeptide-8, acetyl octapeptide-3, dipeptide diaminobutyryl benzylamide diacetate, arginine / lysine peptide, and palmitoyl pentapeptide-4—to obtain an immediate anti-wrinkle peptide composition, thus exhibiting the following beneficial effects: The immediate anti-wrinkle peptide composition of this invention has a good acetylcholine inhibitory effect. After treating mouse hippocampal neurons stimulated with high potassium for 2 hours with the immediate anti-wrinkle peptide composition of this invention, the inhibition rate of acetylcholine reached 43.53-57.49%, which helps to reduce the formation of expression lines and demonstrates a good immediate anti-wrinkle effect. Meanwhile, when the instant anti-wrinkle polypeptide composition of this invention was used to prepare cosmetics, after 2 hours of use by subjects, the number of crow's feet decreased by 14.25-24.49%, the length decreased by 11.35-18.34%, the width decreased by 8.50-14.25%, and the average depth decreased by 7.26-13.00%; the number of nasolabial folds decreased by 16.47-31.67%, the length decreased by 12.84-19.97%, the average width decreased by 8.40-14.56%, and the average depth decreased by 7.38-13.80%. Therefore, the polypeptide composition disclosed in this invention has an instant anti-wrinkle effect and has broad application prospects in anti-wrinkle cosmetics.

[0043] This invention, by further using at least one of 23-hydroxybetulic acid, usnic acid, and modified gallic acid esters as functional additives in the polypeptide composition, has the following beneficial effects: The polypeptide composition disclosed in this invention not only has an immediate anti-wrinkle effect but also effectively improves skin elasticity. After 2 hours of use by subjects, the number, length, average width, and average depth of crow's feet wrinkles were significantly reduced, and the number, length, average width, and average depth of nasolabial folds were significantly improved; at the same time, skin elasticity is improved, resulting in a dual improvement in skin smoothness and plumpness. Therefore, the immediate anti-wrinkle polypeptide composition disclosed in this invention has good immediate anti-wrinkle and skin elasticity-improving effects. Attached Figure Description

[0044] Figure 1 The change rate of the number of crow's feet wrinkles is represented by S1, S2, S3, S4, S5, S6, S7, S8, S9, S10, S11, S8, S9, S10, and S11.

[0045] Figure 2 The change rate of the length of the crow's feet wrinkles is represented by S1, S2, S3, S4, S5, S6, S7, S8, S9, S10, S11, S8, S9, S10, and S11.

[0046] Figure 3 The average width change rate of crow's feet wrinkles is represented by S1, S2, S3, S4, S5, S6, S7, S8, S9, S10, S11, S8, S9, S10, and S11.

[0047] Figure 4 The average depth change rate of crow's feet wrinkles is represented by S1, S2, S3, S4, S5, S6, S7, S8, S9, S10, S11, S8, S9, S10, and S11.

[0048] Figure 5 The change rate of the number of nasolabial folds is given by S1, S2, S3, S4, S5, S6, S7, S8, S9, S10, S11, S8, S9, S10, and S11.

[0049] Figure 6 The change rate of nasolabial fold length is denoted as S1, S2, S3, S4, S5, S6, S7, S8, S9, S10, S11, S8, S9, S10, and S11.

[0050] Figure 7 The average width change rate of nasolabial folds is denoted as S1, S2, S3, S4, S5, S6, S7, S8, S9, S10, S11, S8, S9, S10, and S11.

[0051] Figure 8 The average depth change rate of nasolabial folds is denoted as S1, S2, S3, S4, S5, S6, S7, S8, S9, S10, S11, S8, S9, S10, S11, and S11 are examples of the following: Example 5, Example 6, S3, S4, S5, S6, S10, S11, S12, S13, S14, S15, S16, S17, S18, S19, S10, S11, and S11, respectively.

[0052] Figure 9 This represents the rate of change in skin elasticity.

[0053] Figure 10 The infrared spectrum of the modified gallic acid ester. Detailed Implementation

[0054] The present invention will now be described in further detail with reference to specific embodiments. The given embodiments are merely illustrative of the invention and not intended to limit its scope. The embodiments provided below can serve as a guide for further improvements by those skilled in the art and do not constitute a limitation on the invention in any way.

[0055] Unless otherwise specified, the experimental methods used in the following examples are conventional methods. Unless otherwise specified, the materials and reagents used in the following examples are commercially available.

[0056] Example 1: A polypeptide composition comprising a polypeptide and a preservative, wherein the mass ratio of the polypeptide to the preservative is 1:100. The polypeptide comprises the following components in parts by mass: 30 parts acetyl hexapeptide-8, 20 parts acetyl octapeptide-3, 30 parts dipeptide diaminobutyryl benzylamide diacetate, 10 parts arginine / lysine polypeptide, and 10 parts palmitoyl pentapeptide-4. The preservative comprises the following components in parts by mass: 1 part ethylhexylglycerin and 20 parts 1,2-hexanediol.

[0057] The cosmetic comprises an aqueous phase and an additive phase, with a mass ratio of 1:0.0711. The aqueous phase consists of the following components in parts by mass: 3 parts glycerin, 3 parts butylene glycol, 0.3 parts p-hydroxyacetophenone, 0.5 parts 1,2-hexanediol, 2 parts 1,2-pentanediol, 0.03 parts disodium ethylenediaminetetraacetate (EDTA), and the remainder being deionized water. The additive phase consists of the following components in parts by mass: 5 parts deionized water, 1 part glycerin, 0.1 part 1,2-hexanediol, 0.005 parts PPG-5-cetyl ether-20, 0.005 parts ethylhexylglycerin, and 1 part a polypeptide composition.

[0058] Methods for preparing cosmetics include, S1. Weigh each component according to the mass fractions of butanediol and p-hydroxyacetophenone in the aqueous phase. Mix glycerol, butanediol and p-hydroxyacetophenone, and heat to 65°C while stirring. Stir until the mixture is clear and free of particles to obtain a clear, particle-free liquid for later use.

[0059] S2. Weigh each component according to the mass fractions of glycerol, 1,2-hexanediol, 1,2-pentanediol, disodium EDTA, and deionized water in the aqueous phase. Stir 1,2-hexanediol, 1,2-pentanediol, and disodium EDTA until there are no particles. Then, add 25% of the total volume of deionized water and stir until homogeneous. Add the remaining deionized water and stir until homogeneous to obtain a transparent liquid for later use.

[0060] S3. Add the transparent liquid obtained in step S2 to the transparent, particle-free liquid obtained in step S1, keep it at 65°C for 30 minutes, then cool it to room temperature, add the additive phase and mix to obtain the cosmetic.

[0061] Example 2: A polypeptide composition comprising a polypeptide and a preservative, wherein the mass ratio of the polypeptide to the preservative is 1:100. The polypeptide comprises the following components in parts by mass: 30 parts acetyl hexapeptide-8, 10 parts acetyl octapeptide-3, 30 parts dipeptide diaminobutyryl benzylamide diacetate, 20 parts arginine / lysine polypeptide, and 10 parts palmitoyl pentapeptide-4. The preservative comprises the following components in parts by mass: 1 part ethylhexylglycerin and 20 parts 1,2-hexanediol.

[0062] The cosmetic product is the same as in Example 1, except that the polypeptide composition is replaced with the polypeptide composition of this example.

[0063] The preparation method of the cosmetic is the same as that of Example 1, except that the polypeptide composition is replaced with the polypeptide composition of this example.

[0064] Example 3: A polypeptide composition comprising a polypeptide and a preservative, wherein the mass ratio of the polypeptide to the preservative is 1:100. The polypeptide comprises the following components in parts by mass: 35 parts acetyl hexapeptide-8, 15 parts acetyl octapeptide-3, 20 parts dipeptide diaminobutyryl benzylamide diacetate, 20 parts arginine / lysine polypeptide, and 10 parts palmitoyl pentapeptide-4. The preservative comprises the following components in parts by mass: 1 part ethylhexylglycerin and 20 parts 1,2-hexanediol.

[0065] The cosmetic product is the same as in Example 1, except that the polypeptide composition is replaced with the polypeptide composition of this example.

[0066] The preparation method of the cosmetic is the same as that of Example 1, except that the polypeptide composition is replaced with the polypeptide composition of this example.

[0067] Example 4: A polypeptide composition comprising a polypeptide and a preservative, wherein the mass ratio of the polypeptide to the preservative is 1:100. The polypeptide comprises the following components in parts by mass: 40 parts acetyl hexapeptide-8, 20 parts acetyl octapeptide-3, 20 parts dipeptide diaminobutyryl benzylamide diacetate, 10 parts arginine / lysine polypeptide, and 10 parts palmitoyl pentapeptide-4. The preservative comprises the following components in parts by mass: 1 part ethylhexylglycerin and 20 parts 1,2-hexanediol.

[0068] The cosmetic product is the same as in Example 1, except that the polypeptide composition is replaced with the polypeptide composition of this example.

[0069] The preparation method of the cosmetic is the same as that of Example 1, except that the polypeptide composition is replaced with the polypeptide composition of this example.

[0070] Example 5: A polypeptide composition comprising a polypeptide and a preservative, wherein the mass ratio of the polypeptide to the preservative is 1:100. The polypeptide comprises the following components in parts by mass: 40 parts acetyl hexapeptide-8, 10 parts acetyl octapeptide-3, 30 parts dipeptide diaminobutyryl benzylamide diacetate, 15 parts arginine / lysine polypeptide, and 5 parts palmitoyl pentapeptide-4. The preservative comprises the following components in parts by mass: 1 part ethylhexylglycerin and 20 parts 1,2-hexanediol.

[0071] The cosmetic product is the same as in Example 1, except that the polypeptide composition is replaced with the polypeptide composition of this example.

[0072] The preparation method of the cosmetic is the same as that of Example 1, except that the polypeptide composition is replaced with the polypeptide composition of this example.

[0073] Example 6: A polypeptide composition comprising a polypeptide, a functional additive, and a preservative, wherein the mass ratio of the polypeptide to the preservative is 1:100, and the mass ratio of the polypeptide to the functional additive is 1:0.4. The polypeptide comprises the following components in parts by mass: 30 parts acetyl hexapeptide-8, 20 parts acetyl octapeptide-3, 30 parts dipeptide diaminobutyryl benzylamide diacetate, 10 parts arginine / lysine polypeptide, and 10 parts palmitoyl pentapeptide-4. The preservative comprises the following components in parts by mass: 1 part ethylhexylglycerin and 20 parts 1,2-hexanediol. The functional additive comprises the following components in parts by mass: 5 parts 2,3-hydroxybetulic acid and 10 parts ethanol.

[0074] The cosmetic product is the same as in Example 1, except that the polypeptide composition is replaced with the polypeptide composition of this example.

[0075] The preparation method of the cosmetic is the same as that of Example 1, except that the polypeptide composition is replaced with the polypeptide composition of this example.

[0076] Example 7: The polypeptide composition was the same as in Example 6, except that the mass fraction of 23-hydroxybetulic acid was changed to 2 parts.

[0077] The cosmetic product is the same as in Example 1, except that the polypeptide composition is replaced with the polypeptide composition of this example.

[0078] The preparation method of the cosmetic is the same as that of Example 1, except that the polypeptide composition is replaced with the polypeptide composition of this example.

[0079] Example 8: A polypeptide composition comprising a polypeptide, a functional additive, and a preservative, wherein the mass ratio of polypeptide to preservative is 1:100, and the mass ratio of polypeptide to functional additive is 1:0.4. The polypeptide comprises the following components in parts by mass: 30 parts acetyl hexapeptide-8, 20 parts acetyl octapeptide-3, 30 parts dipeptide diaminobutyryl benzylamide diacetate, 10 parts arginine / lysine polypeptide, and 10 parts palmitoyl pentapeptide-4. The preservative comprises the following components in parts by mass: 1 part ethylhexylglycerin and 20 parts 1,2-hexanediol. The functional additive comprises the following components in parts by mass: 5 parts 23-hydroxybetulic acid, 1.4 parts usnic acid, and 10 parts ethanol.

[0080] The cosmetic product is the same as in Example 1, except that the polypeptide composition is replaced with the polypeptide composition of this example.

[0081] The preparation method of the cosmetic is the same as that of Example 1, except that the polypeptide composition is replaced with the polypeptide composition of this example.

[0082] Example 9: The polypeptide composition was the same as in Example 8, except that the mass fraction of isosine was changed to 0.6 parts.

[0083] The cosmetic product is the same as in Example 1, except that the polypeptide composition is replaced with the polypeptide composition of this example.

[0084] The preparation method of the cosmetic is the same as that of Example 1, except that the polypeptide composition is replaced with the polypeptide composition of this example.

[0085] Example 10: A polypeptide composition comprising a polypeptide, a functional additive, and a preservative, wherein the mass ratio of polypeptide to preservative is 1:100, and the mass ratio of polypeptide to functional additive is 1:0.4. The polypeptide comprises the following components in parts by mass: 30 parts acetyl hexapeptide-8, 20 parts acetyl octapeptide-3, 30 parts dipeptide diaminobutyryl benzylamide diacetate, 10 parts arginine / lysine polypeptide, and 10 parts palmitoyl pentapeptide-4. The preservative comprises the following components in parts by mass: 1 part ethylhexylglycerin and 20 parts 1,2-hexanediol. The functional additive comprises the following components in parts by mass: 5 parts 23-hydroxybetulic acid, 1.4 parts usnic acid, 4.6 parts modified gallic acid ester, and 10 parts ethanol.

[0086] Methods for preparing modified gallic acid esters include, S1. Gallic acid and isobutyric anhydride were mixed and dissolved in N,N-dimethylformamide. 4-Dimethylaminopyridine and triethylamine were added and stirred for 2 hours. The reaction was terminated with dilute hydrochloric acid. The mixture was washed three times with deionized water and filtered to obtain triisobutyrylgalic acid. The molar ratio of gallic acid to isobutyric anhydride was 1:3; the volume ratio of gallic acid to N,N-dimethylformamide was 1 mmol:5 mL; the molar ratio of gallic acid to 4-dimethylaminopyridine was 1:0.1; the molar ratio of gallic acid to triethylamine was 1:5; the concentration of dilute hydrochloric acid was 1 mol / L; the volume ratio of N,N-dimethylformamide to dilute hydrochloric acid was 1:1; and the volume ratio of N,N-dimethylformamide to deionized water was 1:1.

[0087] S2. Triisobutyrylgalic acid and 1,3-dioxane-2-methanol were mixed and dissolved in N,N-dimethylformamide. 4-Dimethylaminopyridine and N,N'-dicyclohexylcarbodiimide were added and stirred for 3 h. The reaction was terminated with dilute hydrochloric acid. The mixture was extracted three times with ethyl acetate, and the organic phase was retained and dried over anhydrous magnesium sulfate. The mixture was purified by vacuum distillation and column chromatography to obtain modified gallic ester. The molar ratio of triisobutyrylgalic acid to 1,3-dioxane-2-methanol is 1:0.5; the volume ratio of triisobutyrylgalic acid to N,N-dimethylformamide is 1 mmol:5 mL; the molar ratio of triisobutyrylgalic acid to 4-dimethylaminopyridine is 1:0.1; the molar ratio of triisobutyrylgalic acid to N,N'-dicyclohexylcarbodiimide is 1:1; the concentration of dilute hydrochloric acid is 1 mol / L; the volume ratio of N,N-dimethylformamide to dilute hydrochloric acid is 1:1; and the volume ratio of N,N-dimethylformamide to ethyl acetate is 1:1.

[0088] The cosmetic product is the same as in Example 1, except that the polypeptide composition is replaced with the polypeptide composition of this example.

[0089] The preparation method of the cosmetic is the same as that of Example 1, except that the polypeptide composition is replaced with the polypeptide composition of this example.

[0090] Example 11: The polypeptide composition was the same as in Example 10, except that the mass fraction of modified gallic acid ester was changed to 3.5 parts.

[0091] The preparation method of the modified gallic acid ester is the same as in Example 10.

[0092] The cosmetic product is the same as in Example 1, except that the polypeptide composition is replaced with the polypeptide composition of this example.

[0093] The preparation method of the cosmetic is the same as that of Example 1, except that the polypeptide composition is replaced with the polypeptide composition of this example.

[0094] Comparative Example 1: The cosmetic product was prepared under the same conditions as Example 1, except that no peptide composition was used.

[0095] The preparation method of the cosmetic is the same as that of Example 1, except that the polypeptide composition was not used.

[0096] Comparative Example 2: A polypeptide composition, comprising acetyl hexapeptide-8.

[0097] The cosmetic product is the same as in Example 1, except that the polypeptide composition is replaced with the polypeptide composition of this example.

[0098] The preparation method of the cosmetic is the same as that of Example 1, except that the polypeptide composition is replaced with the polypeptide composition of this example.

[0099] Comparative Example 3: A polypeptide composition, including acetyl octapeptide-3.

[0100] The cosmetic product is the same as in Example 1, except that the polypeptide composition is replaced with the polypeptide composition of this example.

[0101] The preparation method of the cosmetic is the same as that of Example 1, except that the polypeptide composition is replaced with the polypeptide composition of this example.

[0102] Comparative Example 4: A polypeptide composition comprising dipeptide diaminobutyryl benzylamide diacetate.

[0103] The cosmetic product is the same as in Example 1, except that the polypeptide composition is replaced with the polypeptide composition of this example.

[0104] The preparation method of the cosmetic is the same as that of Example 1, except that the polypeptide composition is replaced with the polypeptide composition of this example.

[0105] Comparative Example 5: A polypeptide composition, including an arginine / lysine polypeptide.

[0106] The cosmetic product is the same as in Example 1, except that the polypeptide composition is replaced with the polypeptide composition of this example.

[0107] The preparation method of the cosmetic is the same as that of Example 1, except that the polypeptide composition is replaced with the polypeptide composition of this example.

[0108] Comparative Example 6: A polypeptide composition, including palmitoyl pentapeptide-4.

[0109] The cosmetic product is the same as in Example 1, except that the polypeptide composition is replaced with the polypeptide composition of this example.

[0110] The preparation method of the cosmetic is the same as that of Example 1, except that the polypeptide composition is replaced with the polypeptide composition of this example.

[0111] Comparative Example 7: The polypeptide composition was the same as in Example 8 except that 23-hydroxybetulic acid was not used.

[0112] The cosmetic product is the same as in Example 1, except that the polypeptide composition is replaced with the polypeptide composition of this example.

[0113] The preparation method of the cosmetic is the same as that of Example 1, except that the polypeptide composition is replaced with the polypeptide composition of this example.

[0114] Comparative Example 8: The polypeptide composition was the same as in Example 10 except that 23-hydroxybetulic acid and mycolic acid were not used.

[0115] The cosmetic product is the same as in Example 1, except that the polypeptide composition is replaced with the polypeptide composition of this example.

[0116] The preparation method of the cosmetic is the same as that of Example 1, except that the polypeptide composition is replaced with the polypeptide composition of this example.

[0117] Comparative Example 9: The polypeptide composition is the same as that in Example 10, except that the modified gallic acid ester is replaced with gallic acid.

[0118] The cosmetic product is the same as in Example 1, except that the polypeptide composition is replaced with the polypeptide composition of this example.

[0119] The preparation method of the cosmetic is the same as that of Example 1, except that the polypeptide composition is replaced with the polypeptide composition of this example.

[0120] Comparative Example 10: The polypeptide composition was the same as in Example 10, except that the modified gallate was replaced with triisobutyrylgalic acid.

[0121] The preparation method of triisobutyrylgallic acid includes, Gallic acid and isobutyric anhydride were mixed and dissolved in N,N-dimethylformamide. 4-Dimethylaminopyridine and triethylamine were added and the mixture was stirred for 2 hours. The reaction was terminated with dilute hydrochloric acid. The mixture was washed three times with deionized water and filtered to obtain triisobutyrylgalic acid. The molar ratio of gallic acid to isobutyric anhydride was 1:3; the volume ratio of gallic acid to N,N-dimethylformamide was 1 mmol:5 mL; the molar ratio of gallic acid to 4-dimethylaminopyridine was 1:0.1; the molar ratio of gallic acid to triethylamine was 1:5; the concentration of dilute hydrochloric acid was 1 mol / L; the volume ratio of N,N-dimethylformamide to dilute hydrochloric acid was 1:1; and the volume ratio of N,N-dimethylformamide to deionized water was 1:1.

[0122] The cosmetic product is the same as in Example 1, except that the polypeptide composition is replaced with the polypeptide composition of this example.

[0123] The preparation method of the cosmetic is the same as that of Example 1, except that the polypeptide composition is replaced with the polypeptide composition of this example.

[0124] Comparative Example 11: The polypeptide composition was the same as in Example 5 except that palmitoyl pentapeptide-4 was not used.

[0125] The cosmetic product is the same as in Example 1, except that the polypeptide composition is replaced with the polypeptide composition of this example.

[0126] The preparation method of the cosmetic is the same as that of Example 1, except that the polypeptide composition is replaced with the polypeptide composition of this example.

[0127] Experimental Example 1: The molecular mechanism underlying expression lines is as follows: During nerve excitation, the body transmits excitation between neurons via synapses. When nerve fiber excitation is transmitted to the synaptic knob in the form of a local current (electrical signal), neuronal vesicles containing acetylcholine (ACh) are stimulated. The neuronal vesicle-associated membrane protein, neuromuscular junction-associated protein, and neuromuscular junction fusion protein form the SNARE complex. The SNARE complex releases the neurotransmitter acetylcholine into the neuromuscular junction, where acetylcholine receptors in the muscle bind to the free acetylcholine, leading to facial muscle contraction. Therefore, the anti-wrinkle effect of the polypeptide composition of this invention can be evaluated by detecting changes in acetylcholine levels in mouse hippocampal neurons.

[0128] The peptide compositions of Examples 1-5, Comparative Examples 2-6, and Comparative Example 11 were mixed with DMEM culture medium to prepare test solutions of the peptide compositions of Examples 1-5, Comparative Examples 2-6, and Comparative Example 11 with a mass concentration of 0.1%. A preservative test solution with a mass concentration of 0.1% was prepared by mixing a preservative with DMEM culture medium. The preservative consisted of the following components in parts by mass: 1 part ethylhexylglycerin and 20 parts 1,2-hexanediol. Mouse hippocampal neurons were then treated with the peptide composition test solution and the preservative test solution, and the acetylcholine content was detected. The specific steps are as follows: Mouse hippocampal neurons were treated with a 1×10⁻⁶ DMEM culture medium. 5 Cells were seeded at a density of 1 / well into 24-well plates, for a total of 14 groups, including a blank control group, a negative control group, a preservative control group, and 11 experimental groups. The blank control group, negative control group, preservative control group, and 11 experimental groups were all incubated at 37°C and 5% carbon dioxide for 24 hours. After incubation, the cells were washed three times with phosphate buffer solution, and then differentiated again at 37°C and 5% carbon dioxide for 24 hours. After differentiation, in the blank control group, 2 mL of DMEM medium was added for 2 h; in the negative control group, 1 mL of DMEM medium and 1 mL of DMEM medium containing 1 mmol / L physostigmine were added for high potassium stimulation for 2 h to determine the basal release; in the preservative control group, 1 mL of preservative test solution and 1 mL of DMEM medium containing 1 mmol / L physostigmine were added for high potassium stimulation for 2 h to eliminate the influence of the preservative; in the 11 experimental groups, 1 mL of the peptide composition test solution of Examples 1-5 and Comparative Examples 2-6 with a mass concentration of 0.1% and 1 mL of DMEM medium containing 1 mmol / L physostigmine were added sequentially for high potassium stimulation for 2 h. After treatment, the supernatant of the negative control group and experimental groups was collected, and the acetylcholine content of each group was determined using an acetylcholine detection kit. The acetylcholine inhibition rate (%) was calculated according to the following formula: Acetylcholine inhibition rate (%) = (Negative control group - Experimental group) / (Negative control group - Blank control group) × 100%.

[0129] There was no significant difference in acetylcholine content between the negative control group and the preservative control group, indicating that ethylhexylglycerin and 1,2-hexanediol, as preservatives, had virtually no inhibitory effect on the release of acetylcholine.

[0130] Table 1 Acetylcholine inhibition rate (%)

[0131] The results are shown in Table 1. Compared with Comparative Examples 2-6, the acetylcholine inhibition rate of Examples 1-5 was significantly increased, with Example 5 showing the best acetylcholine inhibition effect. This indicates that treating mouse hippocampal neurons stimulated with high potassium for 2 hours with the peptide compositions of Examples 1-5 can effectively inhibit the release of acetylcholine, thereby reducing the formation of expression lines and demonstrating a good immediate anti-wrinkle effect.

[0132] In Comparative Example 2, when acetyl hexapeptide-8 was used alone, the acetylcholine inhibition rate was 25.54%. Therefore, when the content of acetyl hexapeptide-8 was 40%, its theoretical acetylcholine inhibition rate should be 10.22%. In Comparative Example 3, when acetyl octapeptide-3 was used alone, the acetylcholine inhibition rate was 12.25%. Therefore, when the content of acetyl octapeptide-3 was 20%, its theoretical acetylcholine inhibition rate should be 2.45%. In Comparative Example 4, when dipeptide diaminobutyryl benzylamide diacetate was used alone... The acetylcholine inhibition rate was 20.64%, therefore, when the content of dipeptide diaminobutyryl benzylamide diacetate was 20%, the theoretical acetylcholine inhibition rate should be 4.13%. In Comparative Example 5, the acetylcholine inhibition rate was 26.15% when arginine / lysine peptide was used alone, therefore, when the content of arginine / lysine peptide was 10%, the theoretical acetylcholine inhibition rate should be 2.06%. In Comparative Example 6, the use of palmitoyl pentapeptide-4 alone did not significantly affect the acetylcholine inhibition rate. Therefore, when 40% acetyl hexapeptide-8, 20% acetyl octapeptide-3, 20% dipeptide diaminobutyryl benzylamide diacetate, 10% arginine / lysine peptide, and 10% palmitoyl pentapeptide-4 were mixed, the theoretical acetylcholine inhibition rate should be 31.11%, which is significantly lower than the acetylcholine inhibition effect of Example 5.

[0133] Compared with Comparative Example 11, the acetylcholine inhibition rates of Examples 1-5 were significantly increased. This indicates that, based on the use of acetyl hexapeptide-8, acetyl octapeptide-3, dipeptide diaminobutyryl benzylamide diacetate, and arginine / lysine peptide, the addition of palmitoyl pentapeptide-4 can further improve the acetylcholine inhibition rate.

[0134] Therefore, compared with using acetyl hexapeptide-8, acetyl octapeptide-3, dipeptide-diaminobutyryl benzylamide diacetate, arginine / lysine peptide, and palmitoyl pentapeptide-4 alone, the combined use of appropriate amounts of these ingredients has the best inhibitory effect on acetylcholine and can achieve good immediate anti-wrinkle effect.

[0135] Experimental Example 2: The cosmetics prepared in Examples 1-11 and Comparative Examples 1-10 were tested for immediate anti-wrinkle efficacy. The specific steps are as follows: S1. Subjects were selected and prepared according to the "Test Method for Seven Efficacy Items of Cosmetics: Anti-wrinkle, Firming, Moisturizing, Oil Control, Repairing, Nourishing, and Soothing" (T / CAB0152-2022). The subjects were 30 women aged 38-55 years whose facial crow's feet and nasolabial folds were assessed by experts to be at levels 2-4. Seven days prior to the test, subjects were required to stop using skincare products of the same type and efficacy as the test sample on the test area. Before the test, subjects were required to cleanse their faces with a fragrance-free, gentle cleanser provided by the laboratory, pat their faces dry with lint-free facial tissues, and wait 30 minutes at 20-22℃ and 40-60% RH for equilibration.

[0136] S2. The test sites were randomly divided into a half-face sample area and a half-face matrix control area. First, the first Antera 3D high-resolution skin photography was performed on the half-face sample area and the half-face matrix control area of ​​the subject, and the first Antera 3D high-resolution skin photography data was obtained, which was denoted as T0.

[0137] S3. Perform half-face testing according to the scheme in Table 2. Apply the cosmetics of Examples 1-11 to the half-face sample area and the cosmetics of Comparative Examples 1-10 to the half-face matrix control area. Two hours after the half-face test, perform a second Antera 3D high-resolution skin photography on both the half-face sample area and the half-face matrix control area to obtain the second Antera 3D high-resolution skin photography data, which is denoted as T1.

[0138] Table 2 Half-face testing protocol

[0139] S4. Multiple quantitative indicators were used to evaluate the change rates of crow's feet and nasolabial folds. The evaluation indicators for crow's feet included quantity, length, average width, and average depth; the evaluation indicators for nasolabial folds included quantity, length, average width, and average depth. In the second hour of treatment, the change rate of each indicator was calculated using the following formula: Change Rate (%) = (T1 - T0) / T0 × 100%. A negative change rate indicates a reduction in the severity of crow's feet or nasolabial folds; a positive change rate indicates an increase in the severity of crow's feet or nasolabial folds.

[0140] Table 3. Change rate (%) of crow's feet wrinkles in the 2nd hour

[0141] Table 3 shows the change rate (%) of crow's feet wrinkles. "*" indicates a significant difference (P < 0.05); "**" indicates an extremely significant difference (P < 0.01); "ns" indicates no significant difference. As shown in Table 3, compared to Comparative Example 1, after treatment with the peptide compositions of Examples 1-5 and Comparative Examples 2-5 for 2 hours, the change rates of the number, length, average width, and average depth of crow's feet wrinkles were all negative and showed varying degrees of reduction. This indicates that after 2 hours of treatment with the cosmetics of Examples 1-5, image analysis showed a significant improvement in the visual appearance of crow's feet wrinkles, with a significant reduction in the number, length, average width, and average depth, demonstrating an immediate anti-wrinkle effect.

[0142] Compared to Comparative Example 1, the changes in the length and average width of crow's feet wrinkles in Comparative Example 3, the length of crow's feet wrinkles in Comparative Example 4, and the number, length, average width, and average depth of crow's feet wrinkles in Comparative Example 6 were not significantly different from those in Comparative Example 1. This indicates that the immediate anti-wrinkle effect of using acetyl octapeptide-3, dipeptide diaminobutyryl benzylamide diacetate, and palmitoyl pentapeptide-4 alone is limited. Meanwhile, compared to Comparative Examples 2-5, the changes in the number, length, average width, and average depth of crow's feet wrinkles in Examples 1-5 all decreased. Among them, Example 5 showed the best improvement in the visual appearance of crow's feet wrinkles through image analysis. This indicates that compared to using acetyl hexapeptide-8, acetyl octapeptide-3, dipeptide diaminobutyryl benzylamide diacetate, arginine / lysine peptide, or palmitoyl pentapeptide-4 alone, using these peptides in appropriate proportions can more effectively achieve an immediate anti-crow's feet effect, especially the combination in Example 5, which showed the best immediate improvement effect on crow's feet wrinkles.

[0143] Figure 1 The percentage change in the number of crow's feet wrinkles. Figure 2 The percentage change in the length of crow's feet wrinkles. Figure 3 This represents the rate of change in the average width of crow's feet wrinkles. Figure 4 This represents the rate of change in the average depth of crow's feet wrinkles. Figures 1 to 4It can be seen that, compared with Example 5, after 2 hours of treatment with the cosmetics of Examples 6-11, the improvement in the number, length, average width, and average depth of crow's feet was significantly enhanced. This is because, in the peptide composition, Examples 6-11 used at least one of 23-hydroxybetulic acid, usnic acid, and modified gallic acid esters as a functional additive, while Example 5 did not. This indicates that the present invention, by introducing at least one of 23-hydroxybetulic acid, usnic acid, and modified gallic acid esters into the peptide composition, can effectively enhance the improvement effect on crow's feet. Compared with Examples 6 and Comparative Examples 7-8, the improvement effect on crow's feet in Examples 8-11 was further enhanced, with Examples 10-11 showing the best effect. This indicates that, in the peptide composition, the best effect is achieved by using these substances in combination in appropriate proportions, rather than using 23-hydroxybetulic acid, usnic acid, and modified gallic acid esters alone.

[0144] Table 4. Change rate (%) of nasolabial folds

[0145] Table 4 shows the change rate (%) of nasolabial folds. "*" indicates a significant difference (P < 0.05); "**" indicates an extremely significant difference (P < 0.01); "ns" indicates no significant difference. As shown in Table 4, compared to Comparative Example 1, the change rates of the number, length, average width, and average depth of nasolabial folds in Examples 1-5 and Comparative Examples 2-5 of this invention were all negative and showed varying degrees of reduction. This indicates that after 2 hours of treatment with the cosmetics from Examples 1-5 of this invention, image analysis showed a significant improvement in the visual appearance of nasolabial folds, with a significant reduction in the number, length, average width, and average depth, demonstrating an anti-wrinkle effect.

[0146] Compared to Comparative Example 1, the change rates of the number, length, and average width of nasolabial folds in Comparative Example 3, the length, average width, and average depth of nasolabial folds in Comparative Example 4, and the number, length, average width, and average depth of nasolabial folds in Comparative Example 6 were not significantly different from those in Comparative Example 1. This indicates that the immediate anti-wrinkle effect of using acetyl octapeptide-3, dipeptide diaminobutyryl benzylamide diacetate, and palmitoyl pentapeptide-4 alone is limited. Meanwhile, compared to Comparative Examples 2-5, the change rates of the number, length, average width, and average depth of nasolabial folds in Examples 1-5 all decreased. Among them, Example 5 showed the best improvement in the visual appearance of nasolabial folds through image analysis. This indicates that compared to using acetyl hexapeptide-8, acetyl octapeptide-3, dipeptide diaminobutyryl benzylamide diacetate, arginine / lysine peptide, or palmitoyl pentapeptide-4 alone, using these peptides in appropriate proportions can more effectively achieve an immediate anti-nasolabial fold effect, especially the combination in Example 5, which showed the best immediate improvement effect on nasolabial folds.

[0147] Figure 5 This represents the rate of change in the number of nasolabial folds. Figure 6 This represents the rate of change in the length of the nasolabial folds. Figure 7 This represents the rate of change in the average width of the nasolabial folds. Figure 8 This represents the rate of change in the average depth of the nasolabial folds. Figures 5 to 8 It can be seen that, compared with Example 5, after 2 hours of treatment with the cosmetics of Examples 6-11, the improvement in the number, length, average width, and average depth of nasolabial folds was significantly enhanced. This is because Examples 6-11 used at least one of 23-hydroxybetulic acid, usnic acid, and modified gallic acid esters as functional additives in the peptide composition, while Example 5 did not. This indicates that the present invention, by introducing at least one of 23-hydroxybetulic acid, usnic acid, and modified gallic acid esters into the peptide composition, can effectively enhance the improvement effect on nasolabial folds. Compared with Examples 6 and Comparative Examples 7-8, the improvement effect on nasolabial folds in Examples 8-11 was further enhanced, with Examples 10-11 showing the best effect. This indicates that, in the peptide composition, the best effect is achieved by using these substances in combination in appropriate proportions, rather than using 23-hydroxybetulic acid, usnic acid, and modified gallic acid esters alone.

[0148] Experimental Example 3: Skin elasticity tests were conducted on the cosmetics prepared in Examples 5-11, Comparative Examples 1, and Comparative Examples 7-10. The specific steps are as follows: S1. Subjects were selected and prepared according to the "Test Method for Seven Efficacy Items of Cosmetics: Anti-wrinkle, Firming, Moisturizing, Oil Control, Repairing, Nourishing, and Soothing" (T / CAB0152-2022). The subjects were 36 women aged 25-35. Seven days prior to the test, subjects were required to stop using skincare products of the same type and efficacy as the test sample on the test area. Before the test, subjects were required to cleanse their faces with a fragrance-free, gentle cleanser provided by the laboratory, pat their faces dry with lint-free facial tissues, and wait 30 minutes at 20-22℃ and 40-60% RH for equilibration.

[0149] S2. The subjects were randomly divided into 12 groups of 3 subjects in each group. A skin elasticity tester was used to test and evaluate the subjects' faces, and the first R² value was obtained. The average value of each group was recorded as R. 21 The R² value is an important parameter for assessing changes in skin elasticity; a higher R² value indicates stronger skin elasticity.

[0150] Subjects in groups S3 and 12 underwent cosmetic treatment for 8 hours in accordance with Examples 5-11, Comparative Examples 1, and Comparative Examples 7-10, respectively. Skin elasticity testing was performed on the subjects' faces to assess their performance, and a second R² value was obtained. The average value of each group was recorded as R. 22 .

[0151] S4. Calculate the skin elasticity change rate (%) using the following formula: Skin elasticity change rate (%) = (R... 22 -R 21 ) / R 21 ×100%.

[0152] Figure 9 This represents the rate of change in skin elasticity. The results are as follows: Figure 9 As shown, compared with Comparative Example 1, the skin elasticity change rates of Examples 5-11 and Comparative Examples 7-10 of the present invention were significantly increased, because Comparative Example 1 did not use the polypeptide composition in its cosmetics. This indicates that cosmetics containing the polypeptide compositions of Examples 5-11 and Comparative Examples 7-10 of the present invention can significantly improve skin elasticity.

[0153] Compared to Example 5, the skin elasticity change rate of Examples 6-11 was significantly increased. This is because Examples 6-11 used at least one of 23-hydroxybetulic acid, usnic acid, and modified gallic acid esters as functional additives in the peptide composition, while Example 5 did not. Compared to Examples 6 and Comparative Examples 7-8, the skin elasticity change rate of Examples 8-11 was significantly increased, with Examples 10-11 showing the best effect. This indicates that in peptide compositions, the optimal effect is achieved by using these substances in combination in appropriate proportions compared to using 23-hydroxybetulic acid, usnic acid, and modified gallic acid esters alone.

[0154] Experiment Example 4: The peptide compositions of Examples 1-11 and Comparative Examples 1-10 were respectively mixed with DMEM culture medium to prepare test solutions of the peptide compositions of Examples 1-11 and Comparative Examples 1-10 at a mass concentration of 0.1%. These solutions were then treated to form fibroblasts, and the relative cell viability was measured using a CCK-8 assay kit to evaluate the cytotoxicity of the peptide compositions of Examples 1-11 and Comparative Examples 1-10. The CCK-8 assay kit was purchased from Shanghai Beyotime Biotechnology Co., Ltd.

[0155] After treatment with the polypeptide compositions of Examples 1-11 and Comparative Examples 1-10 of the present invention, the relative cell viability of fibroblasts was 97.37-100%, which indicates that the polypeptide compositions of Examples 1-11 and Comparative Examples 1-10 of the present invention have low cytotoxicity and good cell safety.

[0156] Example 5: The modified gallic acid ester prepared in Example 10 was characterized by Fourier transform infrared spectroscopy.

[0157] Figure 10 The infrared spectrum of the modified gallic acid ester is shown at 2900 cm⁻¹. -1 An absorption peak for -CH3 appears nearby, at 1750 cm⁻¹.-1 An absorption peak appears at C=O, 1630 cm⁻¹. -1 An absorption peak of C=C appears nearby, at 1100 cm⁻¹. -1 An absorption peak for COC appears. The peak is located at 3400 cm⁻¹. -1 The absence of an absorption peak for -OH in the vicinity indicates that the hydroxyl group of gallic acid has been largely reacted; 1750 cm⁻¹ -1 The presence of an absorption peak at C=O indicates that triisobutyrylgalic acid and 1,3-dioxane-2-methanol have undergone an esterification reaction.

[0158] The conventional operations in the operation steps of this invention are well known to those skilled in the art and will not be described in detail here.

[0159] The embodiments described above provide a detailed explanation of the technical solutions of the present invention. It should be understood that the above descriptions are merely specific embodiments of the present invention and are not intended to limit the present invention. Any changes and modifications made within the scope of the principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A polypeptide composition for instant wrinkle reduction, characterized in that, The product includes a polypeptide and a preservative; the polypeptide includes at least one of acetyl hexapeptide-8, acetyl octapeptide-3, dipeptide diaminobutyryl benzylamide diacetate, arginine / lysine polypeptide, and palmitoyl pentapeptide-4; the polypeptide composition exhibits an acetylcholine inhibition rate of 43.53-57.49% over a 2-hour treatment period.

2. The instant anti-wrinkle polypeptide composition according to claim 1, characterized in that, The polypeptide comprises the following components in parts by weight: 30-50 parts acetyl hexapeptide-8, 1-20 parts acetyl octapeptide-3, 30-50 parts dipeptide diaminobutyryl benzylamide diacetate, 1-20 parts arginine / lysine polypeptide and 0.1-10 parts palmitoyl pentapeptide-4.

3. The instant anti-wrinkle polypeptide composition according to claim 1, characterized in that, The preservative comprises 0.5-2 parts by weight of ethylhexylglycerin and 10-30 parts by weight of 1,2-hexanediol.

4. The instant anti-wrinkle polypeptide composition according to claim 1, characterized in that, The mass ratio of the polypeptide to the preservative is 1:50-200.

5. The instant anti-wrinkle polypeptide composition according to claim 1, characterized in that, The polypeptide composition includes a polypeptide, a functional additive, and a preservative, wherein the functional additive is 23-hydroxybetulic acid.

6. The use of the instant anti-wrinkle polypeptide composition according to any one of claims 1-5 in the preparation of cosmetics.

7. The application according to claim 6, characterized in that, The cosmetic product comprises an aqueous phase and an additive phase.

8. The application according to claim 7, characterized in that, The aqueous phase comprises the following components in parts by weight: 1-5 parts glycerol, 1-5 parts butanediol, 0.1-0.5 parts p-hydroxyacetophenone, 0.1-1 part 1,2-hexanediol, 1-3 parts 1,2-pentanediol, and 0.01-0.05 parts disodium ethylenediaminetetraacetate, with the remainder being deionized water.

9. The application according to claim 7, characterized in that, The additive phase comprises 0.1-5 parts of glycerol, 0.01-1 part of 1,2-hexanediol, 0.001-0.01 parts of PPG-5-cetyl ether-20, 0.001-0.01 parts of ethylhexylglycerol, and 0.05-20 parts by weight of the polypeptide composition of any one of claims 1-5, with the remainder being deionized water.

10. The application according to claim 7, characterized in that, The mass ratio of the aqueous phase to the additive phase is 1:0.01-0.4.

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