Supramolecular palmitoyl pentapeptide-4 and preparation and application thereof

Abstract

The application discloses a supramolecular palmitoyl pentapeptide-4 as well as preparation and application thereof, and belongs to the technical field of supramolecular preparation. According to the application, palmitoyl pentapeptide-4 and small molecular substances (tartaric acid, inositol and ascorbic acid) are stirred at 15-35 DEG C and 100-300 rpm for 5-15 min at a molar ratio of 1-3:1-3, water is then added and uniformly mixed to obtain a mixture, and the mixture is then frozen, water is removed, and grinding is performed to obtain the supramolecular palmitoyl pentapeptide-4. The supramolecular palmitoyl pentapeptide-4 prepared by the application has good water solubility, good stability and high transdermal efficiency.

Description

Technical Field

[0001] This invention relates to a supramolecular palmitoyl pentapeptide-4 and its preparation and application, belonging to the field of supramolecular preparation technology. Background Technology

[0002] The core characteristics of skin aging are the loss of collagen in the dermis and the degradation of the extracellular matrix. Palmitoyl pentapeptide-4 (Pal-KTTKS), as a classic anti-aging signaling peptide, has become a research hotspot and core ingredient in the field of anti-aging skincare because it can mimic type I procollagen fragments and activate the collagen synthesis pathway of fibroblasts. Its lipid solubility is improved through palmitoyl modification, and compared with unmodified pentapeptide (KTTKS), its transdermal permeability and stability are somewhat improved, and it has been widely used in products such as serums and eye creams. However, in current technology, palmitoyl pentapeptide-4 still suffers from three major defects: poor water solubility, insufficient stability, and low transdermal efficiency, which seriously restrict its bioavailability and skincare efficacy.

[0003] Specifically as follows: First, its water solubility is extremely poor, severely limiting its compatibility with formulations. Palmitoyl pentapeptide-4 is a fat-soluble lipopeptide. The strong hydrophobicity of the long palmitoyl chain results in extremely low solubility in aqueous systems, only about 0.1 mg / mL at room temperature, making it difficult to add directly to mainstream water-based formulations such as water-based products and essences. Existing technologies often use organic solvents (ethanol, propylene glycol) or surfactants to aid solubility, which not only irritates the skin and damages the skin barrier but also easily leads to ingredient aggregation and precipitation, affecting product stability and safety. On the other hand, high-concentration oil-phase systems can cause greasiness, poor breathability, and other user experience issues, making it difficult to balance formulation stability, gentleness, and skin feel.

[0004] Secondly, it suffers from insufficient stability and easy degradation and loss of activity. On the one hand, the peptide bonds of palmitoyl pentapeptide-4 are easily affected by temperature, pH, and enzymatic hydrolysis: in environments with pH < 5.5 or pH > 7.0, the peptide bonds are easily hydrolyzed and broken, and the activity retention rate is less than 80% at 50°C. On the other hand, aminopeptidase in the epidermis and dermis rapidly degrades this component. In vitro experiments show that its half-life in skin tissue is only 20-30 minutes, and the degradation rate exceeds 80% after 120 minutes, making it difficult to maintain an effective concentration. Existing technologies improve stability through single stabilizers or pH adjustment, but the effects are limited and cannot simultaneously ensure stability and bioactivity throughout the entire process, resulting in short product shelf life and rapid decline in efficacy.

[0005] Finally, transdermal efficiency is low, making it difficult to reach the dermis for effective action. The lipid bilayer barrier of the stratum corneum strongly blocks large lipopeptides. Palmitoyl pentapeptide-4 has a molecular weight of 802.1 Da, and even after palmitoylation modification, its transdermal efficiency remains extremely low: in vitro transdermal experiments show that only about 0.6% can penetrate into the dermis after 24 hours, with most remaining in the stratum corneum and unable to target fibroblasts. Existing transdermal technologies (such as liposomes and nanoemulsions) suffer from low drug loading capacity, complex preparation, and high cost, and are easily metabolized and cleared by the skin, making it difficult to achieve efficient transdermal penetration and long-term retention, resulting in anti-aging efficacy far below theoretical expectations.

[0006] In summary, although palmitoyl pentapeptide-4-related technologies have achieved basic applications, their poor water solubility, insufficient stability, and low transdermal efficiency are mutually restrictive, becoming key bottlenecks that limit their efficacy and widespread application.

[0007] Supramolecular assembly technology offers a new direction for solving the above problems. By constructing supramolecular aggregates through non-covalent interactions, it is expected to simultaneously improve their water solubility, stability, and transdermal permeability, providing a new strategy for developing highly efficient, stable, and gentle palmitoyl pentapeptide-4-like anti-aging products. Currently, no literature mentions supramolecular palmitoyl pentapeptide-4. Summary of the Invention

[0008] [Technical Issues] Palmitoyl pentapeptide-4 has poor water solubility, insufficient stability, and low transdermal efficiency. No literature has yet mentioned supramolecular palmitoyl pentapeptide-4.

[0009] [Technical Solution] To address the aforementioned problems, this invention provides a supramolecular palmitoyl pentapeptide-4, its preparation, and its application. Specifically, this invention involves mixing palmitoyl pentapeptide-4 and small molecule substances (tartaric acid, inositol, and ascorbic acid) at a molar ratio of 1-3:1-3, stirring at 15-35°C and 100-300 rpm for 5-15 minutes; then adding water and mixing thoroughly to obtain a mixture; finally, freezing the mixture to remove moisture, and grinding to obtain supramolecular palmitoyl pentapeptide-4. The supramolecular palmitoyl pentapeptide-4 prepared by this invention exhibits good water solubility, good stability, and high transdermal efficiency.

[0010] The first objective of this invention is to provide a method for preparing supramolecular palmitoyl pentapeptide-4, comprising the following steps: (1) Mix palmitoyl pentapeptide-4 and small molecule substances at a molar ratio of 1:1-3 at 15-35℃ and 100-300rpm for 5-15 minutes; then add water and mix evenly to obtain a mixture. (2) The mixture was frozen to remove moisture and then ground to obtain supramolecular palmitoyl pentapeptide-4; Among them, the small molecule substances are one or more of tartaric acid, inositol, and ascorbic acid.

[0011] Optionally, in step (1), the ratio of palmitoyl pentapeptide-4 to water is 0.01-0.05 mol: 500-1000 g.

[0012] Optionally, the mixing in step (1) is carried out by stirring at 15-35℃ and 100-500rpm for 20-30 minutes.

[0013] Optionally, in step (2), freezing is performed at -60 to -40°C for 10-30 hours.

[0014] Optionally, the moisture removal in step (2) is achieved by freeze drying, specifically freeze drying at -50 to -30°C for 10-20 hours.

[0015] Optionally, the grinding in step (2) is performed at 20-30℃ and 50-100rpm for 10-15min.

[0016] The second objective of this invention is to prepare supramolecular palmitoyl pentapeptide-4 using the method described herein.

[0017] Optionally, the particle size of supramolecular palmitoyl pentapeptide-4 is 10-20 μm.

[0018] Optionally, supramolecular palmitoyl pentapeptide-4 has good water solubility, good stability, and high transdermal efficiency.

[0019] The third objective of this invention is the application of the supramolecular palmitoyl pentapeptide-4 described herein in the field of cosmetics.

[0020] Optionally, cosmetics include serums, facial oils, face creams, lotions, eye creams, eye serums, face masks, and gel masks.

[0021] The fourth objective of this invention is to provide an essence water in which the supramolecular palmitoyl pentapeptide-4 described in this invention is employed.

[0022] The fifth objective of this invention is to provide a method for improving the water solubility, stability, and transdermal efficiency of palmitoyl pentapeptide-4, which employs the supramolecular palmitoyl pentapeptide-4 described in this invention.

[0023] [Beneficial Effects] (1) The present invention uses palmitoyl pentapeptide-4 and small molecule substances (tartaric acid, inositol, ascorbic acid) as raw materials. The preparation method is simple and can be used on a large scale in industrial applications.

[0024] (2) The supramolecular palmitoyl pentapeptide-4 prepared in this invention significantly improves its physicochemical properties, including water solubility, stability and transdermal performance, without changing the molecular structure of palmitoyl pentapeptide-4, thereby improving bioavailability and playing a synergistic role.

[0025] (3) The supramolecular palmitoyl pentapeptide-4 prepared in this invention has good water solubility, reaching more than 100 mg / mL.

[0026] (4) The supramolecular palmitoyl pentapeptide-4 prepared by the present invention has good stability, is not easily decomposed, and is easy to store and transport; after being stored at 25°C for 7 days, the residual amount of palmitoyl pentapeptide-4 reaches more than 97%; after being stored at 45°C for 7 days, the residual amount of palmitoyl pentapeptide-4 reaches more than 94%.

[0027] (5) The supramolecular palmitoyl pentapeptide-4 prepared by this invention has good transdermal performance, and the cumulative permeation of palmitoyl pentapeptide-4 reaches 44 μg / cm² in 24 hours. 2 above.

[0028] (6) The supramolecular palmitoyl pentapeptide-4 prepared by the present invention has the ability to promote collagen synthesis on the basis of being safe and non-toxic, and the collagen expression level reaches more than 44329. Attached Figure Description

[0029] Figure 1 The infrared spectrum of supramolecular palmitoyl pentapeptide-4 from Example 1 is shown.

[0030] Figure 2 The infrared spectrum of supramolecular palmitoyl pentapeptide-4 in Example 2 is shown.

[0031] Figure 3 The infrared spectrum of supramolecular palmitoyl pentapeptide-4 in Example 3 is shown. Detailed Implementation

[0032] The preferred embodiments of the present invention are described below. It should be understood that the embodiments are for better explanation of the present invention and are not intended to limit the present invention.

[0033] Test method: 1. Solubility test in water: Under constant temperature of 25℃, an excess of supramolecular palmitoyl pentapeptide-4 was added to purified water. After stirring in a sealed container at constant temperature, the mixture was allowed to stand for 24 hours. The saturated supernatant was collected by centrifugation and filtration through a 0.22μm aqueous filter membrane. The absorbance was quantitatively determined at 285nm using ultraviolet spectrophotometry combined with a standard curve.

[0034] Three parallel experiments were set up, and the average value was calculated as its solubility in water.

[0035] 2. High and low temperature cycling test: The supramolecular palmitoyl pentapeptide-4 was prepared as a 5% aqueous solution, frozen at -10°C, and then dissolved at 40°C. This constituted one cycle, which was then repeated. The state after each dissolution was observed to ensure that the liquid remained stable and homogeneous without any solid precipitation. The example used data from 20 cycles.

[0036] 3. Stability test: Supramolecular palmitoyl pentapeptide-4 and palmitoyl pentapeptide-4 were dissolved in deionized water to prepare an aqueous solution with a mass fraction of 0.1% for palmitoyl pentapeptide-4. The samples were stored at 25℃ and 45℃ for 7 days, and the residual amount of palmitoyl pentapeptide-4 was determined at 285nm using a UV spectrophotometer to evaluate its stability.

[0037] 4. Permeability test: In vitro skin penetration testing using a Franz diffusion cell: Undamaged, clean pigskin was placed on a Franz diffusion cell, cuticle side up. Palmitoyl pentapeptide-4 was diluted with deionized water to ensure a palmitoyl pentapeptide-4 concentration of 100 μg / mL in each system. The receiving chamber was filled with physiological saline, and 2 mL of each sample solution was added to the diffusion cell. The mixture was kept at a constant temperature (37 ± 0.1 °C) with continuous stirring. 1 mL of the receiving solution was collected at 1, 2, 4, 6, 12, and 24 hours, and an equal volume of fresh receiving solution (physiological saline) was added. The collected samples were filtered through a 0.45 μm microporous filter and then analyzed by HPLC.

[0038] The cumulative permeation of palmitoyl pentapeptide-4 was calculated using the following formula:

[0039] Among them, Q S Cumulative palmitoyl pentapeptide-4 per unit area (μg / cm²) 2 ), C sn V represents the concentration (mg / mL) of palmitoyl pentapeptide-4 in the receptor fluid measured within the sampling interval. S For the volume of the receptor pool, It is the cumulative palmitoyl pentapeptide-4 concentration in the receptor fluid, S is the sampling volume, and A is the concentration of palmitoyl pentapeptide-4. S It is the effective diffusion area.

[0040] 5. Toxicity and collagen synthesis tests: The effects of palmitoyl pentapeptide-4, the examples, and the comparative examples on cell viability and collagen synthesis in human skin fibroblasts (HDF) were determined through cell experiments.

[0041] After resuscitation, HDF cells were cultured in DMEM medium supplemented with 10% FBS (v / v) and 1% penicillin-streptomycin (v / v). The cells were then incubated at 37°C in a humidified environment with 5% CO2. When cell confluence exceeded 80%, cell passages or subsequent experiments were performed.

[0042] The effect of the sample on HDF cell viability was assessed. HDF cells in the logarithmic growth phase were seeded into 96-well plates at a density of approximately 2 × 10⁶ cells per well. 5 Cells were collected; each well was filled with 100 μL of DMEM medium supplemented with 10% FBS (v / v) and incubated in a CO2 incubator at 37 °C. After 24 h of incubation, the old medium in each well was removed using a pipette (for both cell lines), and 100 μL of fresh medium containing different concentrations of the sample (palmitoyl pentapeptide-4 at 20, 40, 60, 80, and 100 μg / mL) was added to each well as the sample group; the cells were then incubated for another 24 h at 37 °C, with untreated cells as the control group and cell-free fresh medium as the blank group. At the end of the incubation period, 100 μL of MTT solution (0.5 mg / mL, DMEM medium) was added to each well, and incubation was continued at 37 °C for 4 h; after that, the supernatant was removed, and the generated formazan crystals were dissolved in DMSO (gently shaken for 5 min), and the absorbance was measured at 570 nm.

[0043] The formula for calculating cell viability is as follows:

[0044] Among them, A sample A represents the absorbance of the sample solution. blank A represents the absorbance of the blank solution; control The absorbance is the value of the control group solution.

[0045] Cells were cultured using standard methods and seeded into 24-well cell culture plates and incubated for 24 hours. The original culture medium was discarded, and negative control, model control, positive control, and sample groups were established. The negative and model control groups were supplemented with basal maintenance culture medium, while the positive control group was supplemented with DMEM maintenance culture medium containing 0.10% VA. The sample groups were supplemented with DMEM maintenance culture medium containing 0.010% of the control and comparative samples, with three replicates per group. Incubation was continued at 37°C for 24 hours. After incubation, the culture medium in each well was discarded, and the cells were washed twice with PBS. 500 μL of PBS was added to each well. The model control, positive control, and sample groups were then placed in a UV environment with a UVA irradiation intensity of 2.5 mW / cm². 2The irradiation dose was 15 J, and the irradiation was carried out in the dark for 40 minutes. The negative control group was kept in the dark throughout the process and was not exposed to UVA.

[0046] After UVA modeling was completed, the corresponding maintenance culture medium was changed again in each group. The negative control group and the model control group were added with basal maintenance culture medium, the positive control group was replaced with DMEM maintenance culture medium containing 0.10% VA, and the sample group was replaced with DMEM maintenance culture medium containing 0.010% of the samples from each example and comparative example. The cells were incubated at 37°C for 48 h. After the culture was completed, the culture medium in the wells was removed, and the cells were washed once with PBS. The cells were then fixed with 4% paraformaldehyde diluted with PBS for 15 min, and washed thoroughly with PBS 3 times. Subsequently, 0.2% Triton-X-100 diluted with PBS was added for permeabilization at room temperature for 30 min, and then 5% bovine serum albumin (BSA) diluted with PBS was added for blocking at room temperature for 1 h. The primary antibody for type I collagen (Collagen-I) was diluted 1:200 by volume and added to each well. The mixture was incubated overnight at 4 °C. The next day, the secondary antibody was diluted 1:500 by volume and incubated at room temperature in the dark for 2 h. The mixture was then washed three times with PBS. Finally, 10 μL of anti-fluorescence quenching mounting medium containing DAPI fluorescent dye (1.5 μg / mL) was added to each well and the mixture was incubated at room temperature for 10 min. Fluorescence images were observed and acquired using an inverted fluorescence microscope, and the expression level of type I collagen was characterized by the intensity of cellular fluorescence.

[0047] Raw materials used in the examples: Palmitoyl pentapeptide-4: Purchased from Shanghai Yadai Biotechnology Co., Ltd., with a purity of 98%; Tartaric acid: purchased from Shanghai Maclean Biochemical Technology Co., Ltd., with a purity of 99%; Inositol: Purchased from Shanghai Haohong Biomedical Technology Co., Ltd., with a purity of 98%; Ascorbic acid: L-ascorbic acid, purchased from Shanghai Haohong Biomedical Technology Co., Ltd., with a purity of 99.99%.

[0048] Example 1 A method for preparing supramolecular palmitoyl pentapeptide-4 includes the following steps: (1) 0.01 mol palmitoyl pentapeptide-4 and 0.01 mol ascorbic acid were stirred at 25°C and 300 rpm for 10 min; then 600 g of water was added and stirred at 25°C and 300 rpm for 25 min to obtain a mixture; (2) The mixture was frozen at -55℃ for 20h, freeze-dried at -40℃ for 15h to remove moisture, and ground at 25℃ and 100rpm for 10min to obtain supramolecular palmitoyl pentapeptide-4 (particle size 10-20μm).

[0049] The obtained supramolecular palmitoyl pentapeptide-4 was characterized, and the test results are as follows: Figure 1 The image shows the infrared spectrum of supramolecular palmitoyl pentapeptide-4 from Example 1. Figure 1 It can be seen that, compared to palmitoyl pentapeptide-4 and ascorbic acid, the supramolecular palmitoyl pentapeptide-4 of Example 1 exhibits better performance at 3200-3600 cm⁻¹. -1 The OH absorption peaks were broadened; meanwhile, at 1720 cm⁻¹... -1 The absorption peak at the position represents the carbonyl absorption peak. The supramolecular palmitoyl pentapeptide-4 in Example 1 is shifted to a lower wavenumber compared to palmitoyl pentapeptide-4 and ascorbic acid. Therefore, it is speculated that a hydrogen bond is formed between palmitoyl pentapeptide-4 and ascorbic acid; that is, supramolecular palmitoyl pentapeptide-4 is formed.

[0050] Example 2 A method for preparing supramolecular palmitoyl pentapeptide-4 includes the following steps: (1) 0.01 mol palmitoyl pentapeptide-4 and 0.02 mol inositol were stirred at 20°C and 200 rpm for 15 min; then 800 g of water was added and stirred at 25°C and 500 rpm for 20 min to obtain a mixture; (2) The mixture was frozen at -60℃ for 10h, freeze-dried at -50℃ for 10h to remove moisture, and ground at 20℃ and 100rpm for 15min to obtain supramolecular palmitoyl pentapeptide-4 (particle size 10-20μm).

[0051] The obtained supramolecular palmitoyl pentapeptide-4 was characterized, and the test results are as follows: Figure 2 The image shows the infrared spectrum of supramolecular palmitoyl pentapeptide-4 from Example 2. Figure 2 It can be seen that, compared to palmitoyl pentapeptide-4 and inositol, the supramolecular palmitoyl pentapeptide-4 in Example 2 exhibits better performance at 3300-3700 cm⁻¹. -1 The OH absorption peaks were broadened; meanwhile, at 1700 cm⁻¹... -1 The absorption peak at the position represents the carbonyl absorption peak. The supramolecular palmitoyl pentapeptide-4 in Example 2 is shifted to a lower wavenumber compared to palmitoyl pentapeptide-4 and inositol. Therefore, it is speculated that a hydrogen bond is formed between palmitoyl pentapeptide-4 and inositol; that is, supramolecular palmitoyl pentapeptide-4 is formed.

[0052] Example 3 A method for preparing supramolecular palmitoyl pentapeptide-4 includes the following steps: (1) 0.01 mol palmitoyl pentapeptide-4 and 0.03 mol tartaric acid were stirred at 30°C and 200 rpm for 15 min; then 1000 g of water was added and stirred at 25°C and 300 rpm for 20 min to obtain a mixture; (2) The mixture was frozen at -40℃ for 30h, freeze-dried at -30℃ for 20h to remove moisture, and ground at 30℃ and 80rpm for 10min to obtain supramolecular palmitoyl pentapeptide-4 (particle size 10-20μm).

[0053] The obtained supramolecular palmitoyl pentapeptide-4 was characterized, and the test results are as follows: Figure 3 The image shows the infrared spectrum of supramolecular palmitoyl pentapeptide-4 from Example 3. Figure 3 It can be seen that, compared to palmitoyl pentapeptide-4 and tartaric acid, the supramolecular palmitoyl pentapeptide-4 of Example 3 exhibits better performance in the 2800-3600 cm⁻¹ range. -1 The OH absorption peaks were broadened; meanwhile, at 1740 cm⁻¹... -1 The absorption peak at the position represents the carbonyl absorption peak. The supramolecular palmitoyl pentapeptide-4 in Example 3 is shifted to a lower wavenumber compared to palmitoyl pentapeptide-4 and tartaric acid. Therefore, it is speculated that a hydrogen bond is formed between palmitoyl pentapeptide-4 and tartaric acid, that is, supramolecular palmitoyl pentapeptide-4 is formed.

[0054] Example 4 The amount of tartaric acid in Example 3 was adjusted to 0.01 mol, while other aspects remained the same as in Example 3, to obtain supramolecular palmitoyl pentapeptide-4 (particle size 10-20 μm).

[0055] Example 5 The freezing in Example 3 was adjusted to be frozen at -50°C for 30 hours, while other aspects remained the same as in Example 3, to obtain supramolecular palmitoyl pentapeptide-4 (particle size 10-20 μm).

[0056] The supramolecular palmitoyl pentapeptide-4 obtained in Examples 1-5 was subjected to performance testing, and the test results are as follows: Table 1 Performance Testing

[0057] Table 2 Cumulative Permeability

[0058] Comparative Example 1 The amount of tartaric acid in Example 3 was adjusted to 0.05 mol, while other aspects remained the same as in Example 3, and the product was obtained.

[0059] Comparative Example 2 In Example 3, tartaric acid was replaced with nicotinic acid, while other aspects remained the same as in Example 3, and the product was obtained.

[0060] Comparative Example 3 In Example 3, tartaric acid was replaced with glucose, while other aspects remained the same as in Example 3, and the product was obtained.

[0061] Comparative Example 4 A method for preparing supramolecular palmitoyl pentapeptide-4 includes the following steps: (1) Mix 0.01 mol palmitoyl pentapeptide-4, 0.03 mol tartaric acid, and 1000 g water at 30 °C and 200 rpm for 30 min to obtain a mixture; (2) Freeze the mixture at -40℃ for 30h, freeze dry at -30℃ for 20h to remove moisture, and grind at 30℃ and 80rpm for 10min to obtain the product.

[0062] Comparative Example 5 The freezing step (2) in Example 3 is omitted, and the rest is kept the same as in Example 3 to obtain the product.

[0063] Comparative Example 6 In step (2) of Example 3, freeze drying was changed to oven drying (drying at 55°C for 10 hours), while other steps remained the same as in Example 3, and the product was obtained.

[0064] Comparative Example 7 Adjust step (1) of Example 3 as follows: 0.01 mol palmitoyl pentapeptide-4 and 0.03 mol tartaric acid were stirred at 30°C and 200 rpm for 15 min; then 1000 g of water was added and stirred at 55°C and 300 rpm for 20 min to obtain a mixture. Everything else remained the same as in Example 3, and the product was obtained.

[0065] The obtained product was subjected to performance testing, and the test results are as follows: Table 3 Performance Testing

[0066] Example 6 An essence water containing supramolecular palmitoyl pentapeptide-4 prepared in Example 3; The components of the essence water are as follows: Table 4

[0067] The preparation method for essence water is as follows: (1) Add ingredients 1-3 and 5-8 from the formula to the main pot, heat to 70°C, homogenize and disperse evenly, and keep warm for 30 minutes; (2) Start cooling down to 50°C and add ingredient No. 4 into the main pot; (3) Continue to cool down to 40℃, add raw materials No. 9-10, and stir to mix evenly; (4) Cool down to 35°C, discharge the material, and obtain the essence water.

[0068] Comparative Example 8 In Example 6, "supramolecular palmitoyl pentapeptide-4" was adjusted to be palmitoyl pentapeptide-4 and tartaric acid of equal mass, while other components remained the same as in Example 6, resulting in an essence water.

[0069] The obtained essence water was subjected to an anti-aging test, and the test method is as follows: Participants: 30, aged 18-60; Test method: After cleansing their face, the subjects sat quietly for 20 minutes in a constant temperature and humidity waiting area of ​​(21±2)℃ and (50±10)%RH for 14 days. On the day of the test, the subjects did not wear makeup, cleaned their face, and sat quietly for 20 minutes in a constant temperature and humidity waiting area of ​​(21±2)℃ and (50±10)%RH for 14 days for 14 days.

[0070] Test area: face; Testing instrument: Cutometer MPA580 for skin elasticity testing F4; Testing instrument: Visia, used to measure the area of ​​facial wrinkles; Tightness change rate = (Measurement after use - Measurement before use) / Measurement before use × 100%; The lower the value, the better the tightness.

[0071] Wrinkle area change rate = (measured value after use - measured value before use) / measured value before use × 100%. The smaller the value, the less wrinkle area there is.

[0072] The test results are as follows: Table 5

[0073] Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Anyone skilled in the art can make various modifications and alterations without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be determined by the claims.

Claims

1. A method for preparing supramolecular palmitoyl pentapeptide-4, characterized in that, Includes the following steps: (1) Mix palmitoyl pentapeptide-4 and small molecule substances at a molar ratio of 1:1-3 at 15-35℃ and 100-300rpm for 5-15 minutes; then add water and mix evenly to obtain a mixture. (2) The mixture was frozen to remove moisture and then ground to obtain supramolecular palmitoyl pentapeptide-4; Among them, the small molecule substances are one or more of tartaric acid, inositol, and ascorbic acid.

2. The method according to claim 1, characterized in that, In step (1), the ratio of palmitoyl pentapeptide-4 to water is 0.01-0.05 mol: 500-1000 g.

3. The method according to claim 1, characterized in that, In step (1), the mixture is stirred at 15-35℃ and 100-500rpm for 20-30 minutes to achieve uniform mixing.

4. The method according to claim 1, characterized in that, In step (2), freezing is performed at -60 to -40°C for 10-30 hours.

5. The method according to claim 1, characterized in that, In step (2), moisture is removed by freeze drying, specifically freeze drying at -50 to -30°C for 10-20 hours.

6. The method according to claim 1, characterized in that, In step (2), the grinding is carried out at 20-30℃ and 50-100rpm for 10-15min.

7. The supramolecular palmitoyl pentapeptide-4 prepared by the method according to any one of claims 1-6.

8. The application of the supramolecular palmitoyl pentapeptide-4 according to claim 7 in the field of cosmetics.

9. An essence water, characterized in that, The supramolecular palmitoyl pentapeptide-4 described in claim 7 was used.

10. A method for improving the water solubility, stability, and transdermal efficiency of palmitoyl pentapeptide-4, characterized in that, The supramolecular palmitoyl pentapeptide-4 described in claim 7 was used.

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