A pearl-derived polypeptide with multiple skin activities and its applications

Abstract

This invention discloses a pearl-derived polypeptide with multiple skin activities and its applications, relating to the fields of cosmetics and pharmaceuticals. The invention provides a freshwater pearl polypeptide with the amino acid sequence MSLWGQ. This polypeptide exhibits a significant ability to inhibit melanin production, demonstrating a significant melanin-inhibiting effect even at concentrations as low as 0.1 ppm. It also possesses tyrosinase inhibitory activity, making it a potential tyrosinase inhibitor and whitening agent. Furthermore, this pearl polypeptide is non-cytotoxic and does not cause a decrease in cell viability; in fact, it significantly enhances cell viability at a concentration of 10 ppm. This polypeptide also possesses significant anti-wrinkle and skin barrier repair effects, showing broad application prospects in whitening, anti-wrinkle, and skin barrier repair.

Description

Technical Field

[0001] This invention relates to the fields of cosmetics and pharmaceuticals, and more specifically, to a pearl-derived polypeptide with multiple skin activities and its applications. Background Technology

[0002] As a time-honored cosmetic ingredient, pearl's protein components play a crucial role in its efficacy. After enzymatic hydrolysis, these proteins can produce a series of active peptides with different functions. However, there are currently almost no reports that can clearly identify which substance in the pearl protein hydrolysis products brings about its specific skin activity.

[0003] Currently, there are few reports on obtaining skin-active peptides from freshwater pearls, therefore further research is needed on pearl-derived active peptides.

[0004] In view of this, the present invention is proposed. Summary of the Invention

[0005] The purpose of this invention is to provide a pearl-derived polypeptide with multiple skin activities and its applications.

[0006] This invention is implemented as follows:

[0007] In a first aspect, embodiments of the present invention provide the application of a polypeptide in the preparation of a product having at least one of the effects of whitening, anti-wrinkle and skin barrier repair, wherein the amino acid sequence of the polypeptide is shown in SEQ ID NO:1.

[0008] Secondly, embodiments of the present invention provide a composition comprising: a polypeptide and other active ingredients; the amino acid sequence of the polypeptide is shown in SEQ ID NO:1, and the other active ingredients include any one or more of the following: other ingredients with whitening effects, other ingredients with anti-wrinkle effects, and other ingredients with skin barrier repair effects. Thirdly, embodiments of the present invention provide a skin care product comprising: a base for the skin care product and a polypeptide; the amino acid sequence of the polypeptide is shown in SEQ ID NO:1.

[0009] Fourthly, embodiments of the present invention provide the application of the polypeptide in the preparation of a product having melanin-inhibiting activity, wherein the amino acid sequence of the polypeptide is shown in SEQ ID NO:1.

[0010] Fifthly, embodiments of the present invention provide the use of a polypeptide in the preparation of a product having tyrosinase inhibitory activity, wherein the amino acid sequence of the polypeptide is shown in SEQ ID NO:1.

[0011] Sixthly, embodiments of the present invention provide the application of a polypeptide in the preparation of a product with skin barrier repair activity, wherein the amino acid sequence of the polypeptide is shown in SEQ ID NO:1.

[0012] In a seventh aspect, embodiments of the present invention provide the application of a polypeptide in the preparation of a product having anti-wrinkle activity, wherein the amino acid sequence of the polypeptide is shown in SEQ ID NO:1.

[0013] The present invention has the following beneficial effects:

[0014] This invention provides a freshwater pearl polypeptide with the amino acid sequence MSLWGQ. This polypeptide has a significant ability to inhibit melanin production. Even at a concentration as low as one ten-millionth (0.1 ppm), it still has a significant melanin inhibition effect, reducing melanin production by 18.36%. It has tyrosinase inhibitory activity and is a potential tyrosinase inhibitor and whitening agent. At a concentration of 100 ppm, it can inhibit melanin production in zebrafish with an inhibition rate of 23%.

[0015] This pearl polypeptide is non-cytotoxic and does not cause a decrease in cell viability; at a concentration of 10 ppm, it can significantly enhance cell viability.

[0016] This pearl polypeptide can significantly inhibit caudal fin wrinkling caused by photoaging in zebrafish at a concentration of 100 ppm, with an inhibition rate of 56%, demonstrating significant anti-wrinkle effects.

[0017] This pearl polypeptide can significantly promote the proliferation of human immortalized keratinocytes (HaCaT). At concentrations of 11ppm, 33ppm and 100ppm, the proliferation rates are 29.4%, 26.46% and 30.26% respectively, demonstrating its skin barrier repair effect.

[0018] This pearl polypeptide has broad application prospects in whitening, anti-wrinkle and skin barrier repair. Attached Figure Description

[0019] To more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings used in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of the present invention and should not be regarded as a limitation on the scope. For those skilled in the art, other related drawings can be obtained based on these drawings without creative effort.

[0020] Figure 1 The results of mass spectrometry analysis of pearl polypeptide MSLWGQ are shown.

[0021] Figure 2 The diagram shows the results of the binding conformation analysis of the pearl polypeptide MSLWGQ.

[0022] Figure 3 The results of the cytotoxicity assay for the pearl polypeptide MSLWGQ;

[0023] Figure 4 The graph shows the results of melanin production inhibition by the pearl polypeptide MSLWGQ.

[0024] Figure 5 Figure showing the inhibition of melanin production in zebrafish by the pearl polypeptide MSLWGQ.

[0025] Figure 6 Figure showing the effect of pearl polypeptide MSLWGQ on the proliferation of human immortalized keratinocytes;

[0026] Figure 7 Figure showing the results of using the pearl polypeptide MSLWGQ to inhibit photoaging and caudal fin wrinkling in zebrafish. Detailed Implementation

[0027] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer shall apply. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased commercially.

[0028] The triangular sail mussel (Hyriopsis cumingii) is the main species used in freshwater pearl farming, accounting for over 95% of the market. Processed pearl products have long been reported to possess significant skin-beneficial properties, such as whitening, anti-wrinkle, antioxidant effects, and wound-healing promotion.

[0029] On the one hand, embodiments of the present invention provide the application of polypeptides in the preparation of products having at least one of the effects of whitening, anti-wrinkle and skin barrier repair, wherein the amino acid sequence of the polypeptide is shown in SEQ ID NO:1.

[0030] The phrase "having at least one of the effects of whitening, anti-wrinkle and skin barrier repair" in this article specifically means having any one, any two, or all three effects of whitening, anti-wrinkle and skin barrier repair.

[0031] In some embodiments, the product includes any one of cosmetics, skin care products, medical aesthetic products, and pharmaceuticals.

[0032] In some embodiments, the skincare product includes any one of the following: facial cleanser, lotion, toner, moisturizing lotion, eye serum, moisturizing cream, serum, prep product, sunscreen, and mask.

[0033] In some embodiments, the cosmetic includes either a foundation or a makeup base.

[0034] In some embodiments, the medical aesthetic product includes any one of: medical aesthetic facial mask, injectable lyophilized powder, microneedles, and whitening injections.

[0035] In some embodiments, the dosage form of the drug is selected from any one of the following: injection, tablet, capsule, granule, ointment, controlled-release formulation, and aerosol.

[0036] On the other hand, embodiments of the present invention provide a composition comprising: a polypeptide and other active ingredients; the amino acid sequence of the polypeptide is shown in SEQ ID NO:1, and the other active ingredients include any one or more of the following: other ingredients with whitening effects, other ingredients with anti-wrinkle effects, and other ingredients with skin barrier repair effects.

[0037] In some embodiments, the other whitening ingredients include any one or more of arbutin, kojic acid, ascorbic acid, niacinamide, glycyrrhizin, phenylethyl resorcinol, hexapeptide-2, decapeptide-12, and nonapeptide-1.

[0038] In some embodiments, the other anti-wrinkle ingredients include any one or more of the following: Pro-Xylane, retinol, fullerene, resveratrol, ceramide, soy isoflavones, grape seed extract, copper peptide, conopeptide, acetyl hexapeptide-8, acetyl octapeptide-3, and palmitoyl pentapeptide-4.

[0039] In some embodiments, the other ingredients with skin barrier repair effects include any one or more of the following: ceramides, squalane, beta-glucan, asiaticoside, centella asiatica extract, and purslane extract.

[0040] On the other hand, embodiments of the present invention provide a skin product comprising: a base for the skin product and a polypeptide; the amino acid sequence of the polypeptide is shown in SEQ ID NO:1.

[0041] In some embodiments, the skin products include any one of skin care products, cosmetics, and medical aesthetic products.

[0042] In some embodiments, skin care products, cosmetics, and medical aesthetic products are all as described in any of the foregoing embodiments or implementation methods.

[0043] On the other hand, embodiments of the present invention also provide the application of the polypeptide in the preparation of products with melanin-inhibiting activity, wherein the amino acid sequence of the polypeptide is shown in SEQ ID NO:1.

[0044] In some embodiments, the melanin-inhibiting activity includes the ability to inhibit melanin production.

[0045] In some embodiments, the product having melanin-inhibiting activity is specifically a product having melanin-inhibiting activity on cells and / or animals.

[0046] In some embodiments, the cells can be mammalian cells and cell lines. Mammalian cells include cells from any member of the mammalian family, such as human cells, mouse cells, rat cells, monkey cells, and hamster cells. Optionally, the cells are B16 cells.

[0047] In some embodiments, the animals include mammals and humans, specifically including but not limited to mice, rats, monkeys, rabbits, sheep, and humans.

[0048] On the other hand, embodiments of the present invention also provide the application of the polypeptide in the preparation of a product having tyrosinase inhibitory activity, wherein the amino acid sequence of the polypeptide is shown in SEQ ID NO:1.

[0049] In some embodiments, the product having tyrosinase inhibitory activity is specifically a product having tyrosinase inhibitory activity against cells and / or animals. The cells and animals are as described in any of the foregoing embodiments or implementations.

[0050] In some embodiments, the product is the product described in any of the foregoing embodiments or implementations.

[0051] On the other hand, embodiments of the present invention also provide the application of the polypeptide in the preparation of products with skin barrier repair activity, wherein the amino acid sequence of the polypeptide is shown in SEQ ID NO:1.

[0052] In some embodiments, the skin barrier repair activity specifically refers to its ability to promote the proliferation of human and / or animal keratinocytes.

[0053] Furthermore, embodiments of the present invention also provide the application of the polypeptide in the preparation of products with anti-wrinkle activity, wherein the amino acid sequence of the polypeptide is shown in SEQ ID NO:1.

[0054] The product described herein is the same as that described in any of the foregoing embodiments, and will not be repeated here.

[0055] The features and performance of the present invention will be further described in detail below with reference to embodiments.

[0056] Example 1: Identification of Pearl Polypeptide Sequence Structure

[0057] Pearl protein was obtained by acid hydrolysis of submicron pearl powder. This protein was then subjected to neutral protease hydrolysis, ultrafiltration separation, nanofiltration concentration, and lyophilization to obtain a pearl polypeptide composition. The pearl polypeptide composition was then subjected to peptidomics identification and analysis. The identification and analysis methods were as follows: 4 μl of total peptides were separated using a nano-UPLC system (EASY-nLC1200) and data acquisition was performed using a mass spectrometer (Q-Exactive HFX) equipped with a nanoliter ion source. Chromatographic separation was performed using a 100 μm ID × 15 cm reversed-phase column (Reprosil-Pur 120 C18-AQ, 1.9 μm, Dr. Maisch). The mobile phase was an acetonitrile-water-formic acid system, where mobile phase A was 0.1% formic acid-98% aqueous solution (acetonitrile 2%), and phase B was 0.1% formic acid-80% acetonitrile solution (water 20%). After the column was equilibrated with 100% phase A, the sample was directly loaded onto the column via an autosampler and then subjected to column gradient separation at a flow rate of 300 nL / min for 60 min. Mobile phase B ratios were: 2.5% for 2 min, 5%–22% for 34 min, 22%–45% for 20 min, 45%–95% for 2 min, and 95% for 2 min. Mass spectrometry analysis was performed using data-dependent acquisition (DDA) mode, with a total analysis time of 60 min, employing positive ion detection mode. The full scan range was 350–1600 m / z, with a resolution of 120 kJ (@200 m / z), an AGC of 1E6, and a maximum ion implantation time (max IT) of 50 ms. The top 20 ions from the full scan were selected by quadrupole ionization and then fragmented using an HCD for fragment ion scanning. The quadrupole isolation window was 1.2 m / z, the normalized collision energy (NCE) was 30%, the aGC was 1E5, and the max IT was 100 ms. The secondary scan resolution was 15 kJ. Based on peak width, the dynamic exclusion time was set to 30 s; single-charged ions and ions with a valence greater than 6 were not included in the secondary scan. Raw data files were analyzed using Proteome Discoverer software (PD) (version 2.4.0.305, Thermo Fisher Scientific) and its built-in Sequest HT search engine.

[0058] The identification results showed that the pearl polypeptide MSLWGQ (SEQ ID NO:1) was derived from the cytochrome C oxidase subunit II in pearl protein, and the pearl polypeptide MSLWGQ was derived from the first to sixth amino acids of the protein.

[0059] The structural formula of pearl polypeptide is as follows:

[0060] .

[0061] The protein in question is cytochrome C oxidase subunit II, and its amino acid sequence is as follows:

[0062] MSLWGQMGLQEGGSVLGVEVGGLYDYGMFIIVLIFSFVGYLLLSSLVSSFSSRVCLEKQWLEVVWTIVPFFLLLALGLPSIKLLYLMDEINLPESTVKVVGH QWYWSYEYSDSRGSNYSYDSYMVSNPLVAEGYRLLEVDNRCVVANLLQMRGLVTSDDVIHSWAIPSSSIKIDGVPGRVNQVGLCFTRSGVFYGQCSELCGVN HSFMPISVEVVSIKVFSSWVVENHDNVIKKGGDSNQNSKGWSLWGLIVGVAYWVGKGVYFVGKAVIMWHYYLLYYSFYVPGKFLVFSSWDLLQWVVSSGFAL GKWAMWFWGSPGEASLFALHFLAGKFVSGVWFVVTSPVKAAVWAVKGVWSGVVGFISFCGLVFDSVGSSLSSFTDDSFKGFVVSNVSRNTKEFLWTLSHRYS.

[0063] Example 2: Chemical Synthesis of Pearl Polypeptide MSLWGQ

[0064] Chemical synthesis was performed using a PSI319 peptide synthesizer based on the sequence. The specific method is as follows: During preparation, 0.25 mmol of Rink amide-MBHA Resin (degree of substitution 0.68) was swollen in N-methylpyrrolidone (NMP) for 2 hours. 1 mmol of each of the six Fmoc amino acids was dissolved in 4 ml of NMP solution containing 0.275 mol / L diisopropylcarbodiimide (DIC). The dissolved Fmoc amino acids were then sequentially placed at amino acid positions A, B, C, D, E, and F according to the QGLWSM linkage order. The first step involved amino acid coupling. The resin was deprotected using 20% ​​piperidine NMP solvent and then washed with NMP solvent. Subsequently, a DIC / NMP solution containing Fmoc-Gln(Trt)-OH (glutamine Q with a protecting group) at amino acid position A was added to couple the resin for 2 hours. After coupling, the resin was washed with NMP, then deprotected again with 20% piperidine NMP solvent, and the resin-amino acid conjugate was washed with NMP. A DIC / NMP solution containing Fmoc-Gly-OH (glycine G with a protecting group) at amino acid position B was added, and coupling was performed for 2 hours. After coupling, the mixture was washed with NMP, followed by deprotection with 20% piperidine in NMP solvent, and then washed again with NMP. The remaining four amino acids were coupled sequentially using the same method to obtain the peptide-resin coupler. After thoroughly washing the peptide-resin coupler with NMP, it was shrunken with methanol and dried with nitrogen. The peptide-resin coupler was then removed for further cleavage, i.e., shaken in a cleavage reagent (90% trifluoroacetic acid: 4% phenol: 5% triisopropylsilane: 1% water) for 1 hour, followed by filtration. The filtrate was then dried with nitrogen to obtain the crude peptide. After HPLC purification and salt transsalting, a high-purity pearl polypeptide MSLWGQ was obtained. The molecular weight was verified to be correct by mass spectrometry. Figure 1 .

[0065] Example 3: Prediction of Tyrosinase Inhibitory Activity

[0066] Molecular docking simulations showed that the polypeptide (MSLWGQ) possesses tyrosinase inhibitory capabilities. The methods and results are as follows:

[0067] The X-ray crystal structure of 2Y9X was obtained from a protein database. The protonation state of the small molecule was set to pH = 7.4, and the compound was expanded into a 3D structure using Open Babel. A series of preparations were performed on the receptor protein and ligand using the AutoDock tool (ADT3). The docking box was generated using the AutoGrid program, followed by molecular docking using Autodock Vina (1.2.0). The optimal binding conformation was selected to analyze interactions. Finally, a protein-ligand interaction diagram was generated using PyMOL. The 2Y9X protein is represented as a dark blue cartoon model, the ligands as cyan stick models, and their binding sites as magenta stick structures. Nonpolar hydrogen atoms were omitted. Hydrogen bonds, ionic interactions, and hydrophobic interactions are depicted as yellow, magenta, and green dashed lines, respectively. The discovered interactions were analyzed and categorized according to their interactions. Multiple sets of interactions were directly formed between the receptor protein and the small molecule, such as the hydrogen bond formed between Asp25 of 2Y9X and the ligand. Under the influence of these interactions, the binding energy of the protein-small molecule complex is -6.5 kcal / mol. For details on the binding process, see [link to documentation]. Figure 2 .

[0068] Depend on Figure 2 It was found that the pearl polypeptide exhibits multiple interactions with the receptor protein mushroom tyrosinase (2Y9X). Specifically, receptor proteins ASN19, ASN22, and ASP25 interact with the pearl polypeptide via hydrogen bonds, while ASP25 also exhibits hydrophobic interactions. The binding energy between these small proteins is -6.5 kcal / mol. This indicates that the pearl polypeptide possesses tyrosinase inhibitory activity and is a potential tyrosinase inhibitor and whitening agent, which could be applied in cosmetics or pharmaceuticals requiring tyrosinase inhibitors.

[0069] Example 4: Cytotoxicity Experiment

[0070] Cellular experiments demonstrated that the pearl polypeptide (MSLWGQ) was non-cytotoxic and could promote cell proliferation at a concentration of 10 ppm. The methods and results are as follows: B16 cells grown in DMEM medium and in the logarithmic growth phase were injected with (5-6) × 10⁻⁶ cells / cells. 4Seed cells at 100 μl / well in 96-well plates, and add an equal amount of PBS to the wells surrounding the plate to prevent evaporation of the culture medium from affecting the experimental results. Incubate at 37°C with 5% CO2. After 24 h, aspirate the original culture medium with a pipette, wash once with complete culture medium, and add 100 μl of fresh culture medium. Add 10 μl of sample to each well and incubate. After 24 h, discard the culture medium, wash once with PBS, and add 100 μl of culture medium containing 10% CCK8 solution to each well. Incubate at 37°C with 5% CO2 for 1-4 h, observing the color regularly. Once the color deepens, measure the absorbance using a microplate reader. Set dual wavelengths (upper 450 nm, lower 650 nm) and regions. Read reduction→wavelength options: ! lm1-! lm2, display→reduced data.

[0071] The raw data were statistically analyzed using Graphad Prism, yielding the following conclusions: the pearl polypeptide MSLWGQ is non-cytotoxic, does not cause a decrease in cell viability, and significantly enhances cell viability at a concentration of 10 ppm (p < 0.05). (See attached image.) Figure 3 .

[0072] Example 5: Melanin Production Inhibition Experiment

[0073] Experiments on the inhibition of melanin production in B16 cells have demonstrated that pearl polypeptide (MSLWGQ) can effectively inhibit melanin production in B16 cells, with significant effects even at concentrations as low as 0.1 ppm.

[0074] Specifically, the following steps are included:

[0075] B16 cells grown in DMEM medium and in the logarithmic growth phase were subjected to (10-15) × 10⁻⁶ ionization. 4 Cells were seeded at a rate of [number] cells / well in 100 mm cell culture dishes, with 10 mL of culture medium added to each well. The dishes were incubated at 37°C in a 5% CO2 incubator. After 24 h, 100 μL of sample (arbutin 1000 ppm, arbutin 100 ppm, MSLWGQ 100 ppm, MSLWGQ 1 ppm, MSLWGQ 0.1 ppm) was added to each well, with three replicates per concentration. Incubation continued. After 24 h of culture, cells were collected by trypsin digestion, the supernatant was discarded, and the cells were washed twice with PBS, discarding the supernatant again. Cells were lysed in 300 μL of 1N NaOH containing 10% DMSO in each tube and incubated at 80°C for 30 min. 100 μL of cell lysis buffer was added to a 96-well plate, and absorbance was measured at 405 nm using a microplate reader.

[0076] The raw data were statistically analyzed using Graphad Prism, and the results are shown below. Figure 4 The results show that compared with the positive control arbutin, the pearl polypeptide MSLWGQ has a stronger melanin-inhibiting ability. The pearl polypeptide effectively inhibited melanin production in B16 cells, with an inhibition rate of 25.92% at 100 ppm, 23.80% at 1 ppm, and 18.36% at 0.1 ppm. The pearl polypeptide MSLWGQ is a potential skin whitening agent and can be used in the preparation of whitening cosmetics or drugs. Furthermore, this experiment was conducted under normal cell conditions without the addition of any drugs or hormones that promote melanin secretion. Some existing whitening agents on the market are tested using melanin-stimulating models and do not achieve good results in non-melanin-stimulating models. Compared with some melanin-stimulating model tests, this method more accurately reflects the true melanin-inhibiting ability of the samples.

[0077] Example 6: Zebrafish Melanin Inhibition Experiment

[0078] Five zebrafish were randomly selected and placed in each well of a 12-well plate. The sample was administered in water-soluble form, with the concentration of pearl polypeptide MSLWGQ set at 100 ppm. A normal control group was also included, receiving no treatment; the volume per well was 2 mL. The plates were incubated at 28 ℃ in the dark for 45 h. Three zebrafish from each experimental group were randomly selected and photographed under a dissecting microscope. Advanced image processing software was used to analyze and collect data. The melanin signal intensity (S) in the zebrafish head was analyzed, and the whitening effect of the sample was calculated according to a formula to determine whether it possessed whitening efficacy. The image results are shown below. Figure 5 The image processing data is shown in Table 1.

[0079]

[0080] Table 1. Results of data processing for the inhibition of melanin production in zebrafish by the pearl polypeptide MSLWGQ.

[0081]

[0082] The results showed that the pearl polypeptide MSLWGQ at a concentration of 100 ppm could effectively inhibit the production of melanin in zebrafish, with an inhibition rate of 23% (P < 0.05).

[0083] Example 7: Human Immortalized Keratinocyte (HaCaT) Proliferation Experiment

[0084] After the HaCaT cells in the culture dish reached confluence, discard the old culture medium and wash the dish once with PBS. After discarding the PBS, add 1 mL of trypsin and immediately place the dish in a cell culture incubator at 37°C and 5% CO2 concentration for 5 min to digest. Then add 3 mL of 1640 medium (containing 10% serum and 1% penicillin / streptomycin) to stop the digestion. Collect the cells into 15 mL centrifuge tubes and centrifuge at 1000 rpm for 5 min. After centrifugation, discard the supernatant in the centrifuge tubes, add fresh culture medium to suspend the cells, and seed them at a density of 2 × 10⁴ cells / mL, 100 μL / well in a 96-well cell culture plate. The control group is added with an equal volume of cell-free complete culture medium and incubated. After 24 h, add 10 μL of peptide samples (dissolved in PBS) at different concentrations to each well. At this point, the final peptide concentrations are 11, 33, and 100 ppm. Each concentration is used in 5 replicates. The control and blank groups are added with an equal volume of PBS and incubated. After 24 hours, the CCK8 reagent was diluted 1:10 with 1640 medium. The old medium in the 96-well cell culture plate was discarded, and 100 μL of the diluted CCK8 reagent was added to each well. After incubation in an incubator for 1.5 hours, the absorbance was measured at dual wavelengths of 450 nm and 650 nm (reference wavelength) using a microplate reader. The average absorbance of the blank group and the control group was calculated and denoted as Ab and Ac, respectively. Cell viability was calculated using the formula: Cell viability (%) = (Experimental group absorbance - Ab) / (Ac - Ab) × 100%. Statistical analysis was performed using PDSv9.50 software. Significant differences between data were analyzed using the Student's t-test. The results are shown below. Figure 6 .

[0085] The experimental results showed that the pearl polypeptide MSLWGQ could significantly promote the proliferation of HaCaT cells at three concentrations of 11, 33 and 100 ppm, with proliferation rates of 29.40% (P<0.01), 26.46% (P<0.05) and 30.26% (P<0.05), respectively.

[0086] Example 8: Experiment on inhibiting caudal fin wrinkling in a zebrafish photoaging model

[0087] Five zebrafish were randomly selected and placed in each well of a 12-well plate. The sample was administered in water-soluble form, with the concentration of pearl polypeptide MSLWGQ set at 100 ppm. A normal control group and a model control group were also included. Each well contained 2 mL of sample. After incubation at 28 ℃ in the dark for 2 h, except for the normal control group, the zebrafish in each experimental group were irradiated under ultraviolet light three times, each time for 15 min, with a 30 min interval. Incubation continued at 28 ℃ in the dark for another 22 h. Three zebrafish were randomly selected from each experimental group and photographed under a dissecting microscope. Advanced image processing software was used to analyze and collect data. The caudal fin area (A) of the zebrafish was analyzed, and the anti-wrinkle efficacy of the sample was calculated according to the formula to determine whether it possessed anti-wrinkle properties. Image results are shown below. Figure 7 The data processing results are shown in Table 2.

[0088]

[0089] Table 2. Data processing results of the pearl peptide MSLWGQ inhibiting photoaging and caudal fin wrinkling in zebrafish.

[0090]

[0091] The results showed that the pearl polypeptide MSLWGQ at a concentration of 100 ppm could effectively inhibit the shrinkage of the tail fin of zebrafish caused by photoaging, and had an anti-wrinkle effect with an anti-wrinkle rate of 56% (P<0.01).

[0092] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. The application of polypeptides in the preparation of products with at least one of the following effects: whitening, anti-wrinkle, and skin barrier repair, characterized in that, The amino acid sequence of the polypeptide is shown in SEQ ID NO:

1.

2. The application according to claim 1, characterized in that, The products include any one of the following: cosmetics, medical aesthetic products, and pharmaceuticals.

3. The application according to claim 2, characterized in that, The products include skincare products.

4. The application according to claim 3, characterized in that, The skincare products include any one of the following: facial cleanser, lotion, toner, moisturizer, serum, skin primer, sunscreen, and mask.

5. The application according to claim 4, characterized in that, The skincare products include moisturizing lotions.

6. The application according to claim 4, characterized in that, The skincare products include an eye serum.

7. The application according to claim 2, characterized in that, The cosmetics include either foundation or sunscreen.

8. The application according to claim 2, characterized in that, The medical aesthetic products include any one of the following: medical aesthetic masks, injectable freeze-dried powder, microneedles, and whitening injections.

9. The application according to claim 2, characterized in that, The dosage form of the drug is selected from any one of the following: injection, tablet, capsule, granule, ointment, controlled release, and aerosol.

10. A composition, characterized in that, It includes: polypeptides and other active ingredients; the amino acid sequence of the polypeptide is shown in SEQ ID NO:1; The other active ingredients include any one or more of the following: other ingredients with whitening effects, other ingredients with anti-wrinkle effects, and other ingredients with skin barrier repair effects.

11. The composition according to claim 10, characterized in that, Other ingredients with whitening effects include any one or more of the following: arbutin, kojic acid, ascorbic acid, niacinamide, glycyrrhizin, phenylethyl resorcinol, hexapeptide-2, decapeptide-12, and nonapeptide-1.

12. The composition according to claim 10, characterized in that, Other anti-wrinkle ingredients include any one or more of the following: Pro-Xylane, retinol, fullerene, resveratrol, ceramide, soy isoflavones, grape seed extract, copper peptide, conopeptide, acetyl hexapeptide-8, acetyl octapeptide-3, and palmitoyl pentapeptide-4.

13. The composition according to claim 10, characterized in that, Other ingredients with skin barrier repair effects include any one or more of the following: ceramides, squalane, beta-glucan, centella asiatica extract, and purslane extract.

14. The composition according to claim 13, characterized in that, Other ingredients with skin barrier repair effects include: Centella asiatica extract.

15. A skin care product, characterized in that, It includes: The base and peptides of skin care products; The amino acid sequence of the polypeptide is shown in SEQ ID NO:

1.

16. The skin product according to claim 15, characterized in that, The skin care products include any one of cosmetics and medical aesthetic products.

17. The skin product according to claim 16, characterized in that, The base of cosmetics includes either liquid foundation or primer.

18. The skin product according to claim 16, characterized in that, The skin products mentioned include skin care products.

19. The skin product according to claim 18, characterized in that, The base of the skincare product includes any one of the following: facial cleanser, lotion, toner, moisturizer, serum, skin primer, sunscreen, and mask.

20. The skin product according to claim 19, characterized in that, The base of the skincare product includes a moisturizing lotion.

21. The skin care product according to claim 19, characterized in that, The skincare products include an eye serum.

22. The skin product according to claim 16, characterized in that, The medical aesthetic products include any one of the following: medical aesthetic masks, injectable freeze-dried powder, microneedles, and whitening injections.

23. The application of polypeptides in the preparation of products with melanin-inhibiting activity, characterized in that, The amino acid sequence of the polypeptide is shown in SEQ ID NO:

1.

24. The application according to claim 23, characterized in that, The melanin-inhibiting activity includes the ability to inhibit melanin production.

25. The application of polypeptides in the preparation of products with tyrosinase inhibitory activity, characterized in that, The amino acid sequence of the polypeptide is shown in SEQ ID NO:

1.

26. The application of polypeptides in the preparation of products with skin barrier repair activity, characterized in that, The amino acid sequence of the polypeptide is shown in SEQ ID NO:

1.

27. The application according to claim 26, characterized in that, The skin barrier repair activity includes the ability to promote the proliferation of human immortalized keratinocytes.

28. The application of polypeptides in the preparation of products with anti-wrinkle activity, characterized in that, The amino acid sequence of the polypeptide is shown in SEQ ID NO:

1.

29. The application according to any one of claims 23 to 28, characterized in that, The product is the product described in any one of claims 1 to 9.

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