A class of peptides that can scavenge reactive oxygen species and increase SIRT gene expression and their applications

Abstract

The present invention belongs to the field of biological polypeptide research technology, and specifically relates to a class of polypeptides and applications thereof that can remove reactive oxygen and increase SIRT gene expression. The general formula of the polypeptide sequence is nicotinic acid-C-C-A-r-B-r-B-r-B-r-A-R-G-D-A-NH2; wherein G is glycine, R is arginine, D is aspartic acid, and r is arginine; A is one of tryptophan, tyrosine, and phenylalanine, B is one of tryptophan, phenylalanine, tyrosine, and 3-cyclohexylalanine, C is one of tryptophan, tyrosine, and phenylalanine, A, B, and C are different from each other, r is D configuration, and the remaining amino acids are L configuration. The polypeptide of the present invention can significantly increase the expression of longevity protein-related genes in HaCaT cells, and can effectively remove ROS in cells, so it has broad application prospects in anti-aging and anti-oxidation.

Description

Technical Field

[0001] The present invention belongs to the technical field of biological polypeptide research, and specifically relates to a class of polypeptides capable of removing reactive oxygen species and increasing SIRT gene expression and applications thereof. Background Art

[0002] Mitochondria are the main place where cells produce reactive oxygen species (ROS). Under normal conditions, intracellular ROS can act as intracellular signal molecules to regulate the body's normal physiological functions. When tissues are damaged, the reactive oxygen species (ROS) in mitochondria increase greatly, thereby damaging mitochondria and causing mitochondrial dysfunction. Antioxidants can improve damage by reducing oxidative damage to mitochondria.

[0003] During the aerobic metabolism in mitochondria, 1% to 2% of the total oxygen consumption is converted into oxygen free radicals, and the generation of ROS increases with age and environmental exposure. The large amount of ROS produced first leads to mitochondrial dysfunction, morphological changes and mitochondrial DNA (mtDNA) damage. mtDNA is located in the mitochondrial matrix and is more susceptible to ROS damage than nuclear DNA.

[0004] This vicious cycle proceeds in an age-dependent manner, leading to a gradual decline in the biological functions of tissue cells and accelerating the aging process; mitochondrial metabolic disorders are closely related to cell lifespan and overall skin aging, from mitochondrial genomic DNA (mtDNA) damage to reduced mitochondrial biosynthesis (PGC-1α), to cell aging centered on ATP depletion (keratinocyte aging, melanocyte aging, fibroblast aging). The curtain is drawn, causing overall skin aging in a ripple effect.

[0005] Sirtuins (SIRT) are a class of proteins closely related to cell metabolism, growth and aging, encoded by SIRT genes; SIRT has 7 subtypes, SIRT1-SIRT7, and is a class III histone deacetylase that is widely present in living organisms and depends on nicotinamide adenine dinucleotide (NAD+). It participates in regulating many important biological processes, including maintaining genome stability, regulating metabolism and regulating stem cells. SIRT proteins play an important role in cell stress resistance, energy metabolism, cell apoptosis and aging.

[0006] As a longevity protein, SIRT naturally plays an important role in skin anti-aging. SIRT is involved in a variety of important skin-related cellular functions and processes, including aging, UV damage response, oxidative stress and wound repair.

[0007] SIRT1: SIRT1, which is localized in the cell nucleus, mainly plays a role in promoting differentiation in skin keratinocytes, which is a key link in maintaining the integrity of the skin barrier.

[0008] SIRT2: Located in the cell nucleus and cytoplasm, it plays a role in reducing oxidative stress; decreased SIRT2 activity may cause signs of aging in skin cells and reduce their natural mechanical properties that support the skin.

[0009] SIRT3: Located in mitochondria, it has been found to regulate microtubules, which are components of the cytoskeleton and play an important role in cell shape and structure. As we age, SIRT3 in skin cells becomes very sensitive to the external environment, and ultraviolet radiation can reduce SIRT3 expression.

[0010] SIRT6: Located in the cell nucleus, it can inhibit the nuclear transcription factor NF-κB, thereby downregulating the inflammatory response.

[0011] Given the negative effects of excessive ROS and reduced SIRT expression, it is necessary to develop an agent that can simultaneously reduce ROS levels and increase SIRT expression. Summary of the invention

[0012] The purpose of the present invention is to provide a class of polypeptides that can remove reactive oxygen and increase the expression of SIRT genes and their applications, so as to solve the above-mentioned technical problems.

[0013] The present invention adopts the following technical solution:

[0014] In the first aspect, the present invention provides a class of polypeptides that can scavenge reactive oxygen and increase the expression of SIRT genes, the general formula of the polypeptide sequence is nicotinic acid-CCArBrBrBrARGDA-NH2; wherein G is glycine, R is arginine, D is aspartic acid, r is arginine; A is one of tryptophan, tyrosine, and phenylalanine, B is one of tryptophan, phenylalanine, tyrosine, and 3-cyclohexylalanine, C is one of tryptophan, tyrosine, and phenylalanine, A, B, and C are different from each other, and in the polypeptide, r is in D configuration, and the remaining amino acids are all in L configuration.

[0015] The polypeptide provided by the present invention has the functions of both scavenging free radicals and increasing the expression of SIRT protein genes. Preferably, when A is tyrosine, B is tryptophan, and C is phenylalanine, the comprehensive function is the strongest and can be used to prepare free radical scavenging and / or anti-aging products.

[0016] In a second aspect, the present invention provides use of the polypeptide in preparing free radical scavenging cosmetics or medicines.

[0017] In a third aspect, the present invention provides an application in preparing cosmetics or medicines for resisting aging of human skin cells.

[0018] In a fourth aspect, the present invention provides a drug containing the polypeptide, wherein the polypeptide exists in the form of acetate.

[0019] Furthermore, the medicine also contains pharmaceutically acceptable excipients.

[0020] Furthermore, the auxiliary material is an auxiliary material required for preparing an aqueous extract, a powder, a lotion, a tincture, an oil, an emulsion, an ointment, a plaster or an aerosol.

[0021] In a fourth aspect, the present invention provides a cosmetic containing the polypeptide.

[0022] Furthermore, the cosmetic is one of essence, toner, lotion or cream.

[0023] The present invention has the following beneficial effects:

[0024] The polypeptide provided by the present invention can significantly increase the expression of three longevity protein-related genes, SIRT1, SIRT3, and SIRT6, in HaCaT cells, and can effectively remove ROS in cells. Therefore, it has broad application prospects in anti-aging and anti-oxidation, and can be used to prepare cosmetics or drugs for free radical scavenging and anti-aging of human skin cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 This is the MS image of the targeting peptide.

[0026] Figure 2 HPLC chart of the targeting peptide.

[0027] Figure 3 The cell viability of HaCat after being acted upon by targeting peptides at different dilution multiples.

[0028] Figure 4 The gene expression levels of SIRT1, SIRT3 and SIRT6 in HaCat after the action of targeting peptides at different dilution multiples.

[0029] Figure 5 The ROS levels in HaCat after the action of targeting peptides at different dilution multiples. DETAILED DESCRIPTION

[0030] The present invention is described in detail below in conjunction with the accompanying drawings and specific examples, but should not be construed as limiting the present invention. Unless otherwise specified, the technical means used in the following examples are conventional means well known to those skilled in the art, and the materials, reagents, etc. used in the following examples, unless otherwise specified, can be obtained from commercial sources.

[0031] The present invention provides a class of polypeptides that can remove reactive oxygen and increase the expression of SIRT genes. The general formula of the polypeptide sequence is nicotinic acid-CCArBrBrBrARGDA-NH2; wherein G is glycine, R is arginine, D is aspartic acid, and r is arginine; A is one of tryptophan, tyrosine, and phenylalanine, B is one of tryptophan, phenylalanine, tyrosine, and 3-cyclohexylalanine, and C is one of tryptophan, tyrosine, and phenylalanine. A, B, and C are different from each other. In the polypeptide, r is in D configuration, and the remaining amino acids are all in L configuration.

[0032] The following examples take a polypeptide (hereinafter referred to as a targeting peptide) in which A is tyrosine, B is tryptophan, and C is phenylalanine as an example to verify its effect on removing reactive oxygen species and increasing the expression of SIRT genes.

[0033] Example 1: Preparation and characterization of targeting peptides

[0034] The peptide compound was synthesized by solid phase method, using Fmoc protected amino acid as raw material, Rink Amide MBHAResin (substitution degree of 0.8mmol / g) as solid phase carrier, DIEA / HBTU as condensation agent, and finally the peptide was cleaved from the resin with a mixed solution of trifluoroacetic acid / triisopropylsilane, and the crude peptide was obtained by crystallization and drying with methyl tert-butyl ether. After the crude peptide was dissolved, it was purified and prepared by reverse high performance liquid chromatography (C18, 10um, 100A filler) for multiple times, and the gradient elution segmented sampling method was adopted. Through high performance liquid chromatography interval detection and impurity analysis, a purified intermediate with a purity of more than 90% was obtained, and finally 32g\L ammonium acetate was used for salt conversion. After the final detection was correct, the organic solvent was replaced by a nanofiltration device to reduce its residue and achieve the purpose of enrichment, and it was lyophilized to obtain 0.5g of the lyophilized powder of the targeting peptide.

[0035] The targeting peptide was analyzed by electrospray mass spectrometry, and the molecular weight (e.g. Figure 1 , Figure 2 The molecular weight is basically consistent with the theoretical molecular weight.

[0036] Example 2: Cytotoxicity assay of targeting peptides

[0037] 1. Purpose of testing

[0038] The safe concentration of the sample acting on HaCat cells was detected and the cytotoxicity of the sample was evaluated.

[0039] 2. Detection basis

[0040] Refer to GB / T 16886.5-2017 / 180 10993-5:2009 "Biological Evaluation of Medical Devices" Part 5: In vitro cytotoxicity test.

[0041] 3 Reagents or materials

[0042] 3.1 Cells

[0043] Human immortalized keratinocytes: HaCat.

[0044] 3.2 Reagents and materials

[0045] MTT powder (Sigma), high glucose DMEM (Gibco), PBS powder (XINTAI), Trypsin (Gibco), disodium ethylenediaminetetraacetic acid / EDTA (Sinopharm), fetal bovine serum / FBS (Sijiqing), dimethyl sulfoxide / DMSO (Sinopharm), TryPsin (Gibco), double distilled water, 96-well plate.

[0046] 4. Instruments and Equipment

[0047] Microplate reader (Bio-tek), electronic balance (Pioneer), vortexer (Hangzhou Miou), pipette (Eppendorf), carbon dioxide cell culture incubator (Thermo), cell counter (Invitrogen), inverted fluorescence microscope (Nikon), oscillator (Kylin-Bell), clean bench (Sujing Antai).

[0048] 5. Detection Methods

[0049] 5.1 Sample preparation

[0050] The target peptide mother solution with a concentration of 2 mg / mL was diluted with deionized water in sequence (10 times, 20 times...1280 times) to prepare sample solutions with the required dilution multiples, which were recorded as sample 1, sample 2...sample 8 in sequence.

[0051] 5.2 Test steps

[0052] Cytotoxicity test: MTT method was used to detect cytotoxicity. HaCat cells in logarithmic growth phase were cultured at 1.5×10 5 Cells / mL were inoculated into a 96-well plate, 100 μL / well, and cultured for 24 hours. When the cell plating rate in the 96-well plate reached 40% to 60%, the drug was administered. ① Sample group: 200 μL of culture medium containing samples of different concentrations was added to each well; ② Control group (SC): 200 μL of culture medium was added; ③ Blank group (BC): no cells, only 200 μL of culture medium was added; cultured for 24 hours. The discarded liquid was replaced with culture medium containing MTT (0.5 mg / mL), 200 μL / well, and culture was continued for 4 hours. 150 μL / well of DMSO was added to the discarded liquid, and after sufficient oscillation for color development, the OD was detected using an enzyme reader. 490 , the relative cell viability was calculated according to formula (1):

[0053]

[0054] Cytotoxicity criteria: If the cell survival rate is greater than 70%, it is considered to be free of cytotoxicity; otherwise, it is considered to be cytotoxic.

[0055] 5.3 Statistics

[0056] The software was used to perform a T-Test on the values ​​of the sample group and the SC group. P < 0.05 was considered to be statistically significant compared with the SC group, indicated by *, and P < 0.01 was considered to be extremely statistically significant compared with the SC group, indicated by **.

[0057] 6 Test results

[0058] 6.1 Cytotoxicity test results

[0059] The results of the cytotoxicity test are shown in Table 1 and the resulting bar graph is shown in Figure 3 ,The results showed that 2 mg / mL of the targeting peptide had no obvious cytotoxicity when diluted 40 times.

[0060] Table 1 Targeting peptide (2 mg / mL) cytotoxicity results

[0061] sample Sample 1 Sample 2 Sample 3 Sample 4 Sample 5 Sample 6 Sample 7 Sample 8 SC Concentration (times) 10 20 40 80 160 320 640 1280 / Mean(%) 2.69 42.63 91.02 90.92 90.71 93.78 98.06 100.44 100.00 SD 0.015 0.029 0.028 0.013 0.030 0.019 0.019 0.017 0.013 P 0.000 0.000 0.007 0.001 0.008 0.010 0.215 0.746 1.000

[0062] “ / ” means the item is not included.

[0063] Example 3: Fluorescence quantitative PCR detection of targeted peptides

[0064] 1. Purpose of testing

[0065] The test samples were applied to HaCaT cells, and the changes in gene levels were detected by real-time fluorescence quantitative PCR to evaluate the regulatory effects of the samples at the gene level.

[0066] 2. Detection basis

[0067] Refer to GB / T 16886.5-2017 "Biological Evaluation of Medical Devices".

[0068] 3 Reagents or materials

[0069] 3.1 Cells

[0070] Human keratinocytes: HaCaT cells.

[0071] 3.2 Reagents and materials

[0072] High-glucose DMEM (Gibco), PBS powder (XINTAI), Trypsin (Gibco), disodium ethylenediaminetetraacetic acid / EDTA (Sinopharm), fetal bovine serum / FBS (Sijiqing), TryPsin (Gibco), RNAiS OPLUS (TaKaRa), chloroform (Sinopharm), isopropanol (Sinopharm), 75% alcohol, DEPC water, double distilled water, steel ruler, marker pen, 6-well plate.

[0073] RNAiso Plus (TaKaRa), Evo M-MLV reverse transcription reagent premix (Aicore Biotechnology), SYBR GreenPro Taq HS premix qPCR kit (Aicore Biotechnology).

[0074] 4. Instruments and Equipment

[0075] Electronic balance (Pioneer), spiral mixer (Hangzhou Miou), pipette (Eppendorf), carbon dioxide incubator (Thermo), cell counter (Invitrogen), inverted fluorescence microscope (Nikon), oscillator (Kylin-Bell), clean bench (Sujing Antai), PCR instrument (Roche, Lightcycler96), ultra-micro spectrophotometer (DENOVIX), low-temperature centrifuge (Eppendorf), water bath (Kewei).

[0076] 5. Detection Methods

[0077] 5.1 Cell culture

[0078] After the HaCaT cells stored in liquid nitrogen are revived, they are routinely cultured in high-glucose DMEM complete culture medium containing 10% FBS at 37°C, 5% CO2, and 95% relative humidity. They need to be subcultured 2 to 3 times before the experiment. An inverted microscope is used to observe the cell morphology, growth status, contamination, etc. When the cell fusion rate reaches about 80%, subculture can be performed.

[0079] 5.2 Sample Information

[0080] The target peptide stock solution with a concentration of 2 mg / mL was diluted 80 times and 1280 times to prepare sample solutions with the required dilution multiples.

[0081] 5.3 Test steps

[0082] Gene level detection: Real-time fluorescence quantitative PCR method was used to culture 5×10 HaCaT cells in logarithmic growth phase. 5The cells were inoculated into a 6-well plate at a density of cells / well and incubated overnight in an incubator (37°C, 5% CO2). When the cell plating rate in the 6-well plate reached 40% to 50%, the targeted peptide (80 times, 1280 times) was administered, and the dosage per well was 2 mL. After the administration, the 6-well plate was placed in an incubator (37°C, 5% CO2) and incubated for 24 hours. After the incubation, 2 mL / well PBS was used for washing 3 times, 1 mL RNAiso Plus was added to each well, and the cells were lysed by blowing. The cell lysate was collected for RNA extraction, reverse transcription, and fluorescence quantitative PCR to detect gene levels. 2 -△△CT Method to calculate the results.

[0083] 5.4 Statistics

[0084] The software was used to perform T-Test on the values ​​of the sample group and the SC group. P < 0.05 was considered to be statistically significant compared with the SC group, indicated by "*", and P < 0.01 was considered to be extremely statistically significant compared with the SC group, indicated by "**".

[0085] 6 Test results

[0086] 6.1 Gene level detection

[0087] Fluorescence quantitative PCR was used to detect SIRT1, SIRT3, and SIRT6 related genes. The results are shown in Table 2 and the generated bar graph is shown in Figure 4 The results showed that after the sample targeting peptide acted on HaCaT cells for 24 hours, the sample was diluted 80 times and 1280 times, and the SIRT1, SIRT3, and SIRT6 genes were significantly increased.

[0088] Table 2 Results of fluorescent quantitative PCR gene detection

[0089]

[0090]

[0091] Example 4: Reactive oxygen species (ROS) inhibition assay of targeted peptides

[0092] 1. Purpose and principle of detection

[0093] 1.1 Purpose

[0094] The firming and anti-wrinkle effects of the samples were evaluated based on the reactive oxygen species (ROS) inhibition test of human keratinocytes.

[0095] 1.2 Principle

[0096] Cellular reactive oxygen species detection: (DCFH-DA) has no spontaneous fluorescence and can freely pass through the cell membrane. After entering the cell, it is hydrolyzed by esterase to generate DCFH, which cannot penetrate the cell membrane and accumulates in the cell. The reactive oxygen species in the cell can oxidize the non-fluorescent DCFH to generate fluorescent DCF. The green fluorescence intensity is proportional to the level of reactive oxygen species in the cell. Hydrogen peroxide (H2O2), ultraviolet rays, blue light, etc., as exogenous oxidants, can induce an increase in the ROS content in human keratinocytes. The ROS inhibition rate is calculated by comparing the changes in the fluorescence intensity of the intracellular fluorescent dye DCFH-DA after administration of the negative control and the test substance.

[0097] 2. Detection basis

[0098] Refer to T / SHRH cosmetics firming and anti-wrinkle efficacy test - in vitro keratinocyte reactive oxygen species (ROS) inhibition test method.

[0099] 3 Reagents or materials

[0100] 3.1 Cells

[0101] Human keratinocytes: HaCaT, purchased from Guangdong Boxi Biological Co., Ltd.

[0102] 3.2 Reagents and materials

[0103] High-glucose DMEM (Gibco), PBS powder (XINTAI), Trypsin (Gibco), disodium ethylenediaminetetraacetic acid / EDTA (Sinopharm), fetal bovine serum / FBS (Sijiqing), Trypsin (Gibco), marker pen, and 6-well plate.

[0104] 250μM hydrogen peroxide / H2O2 (Shanghai test), reactive oxygen species ROS kit (Biyuntian).

[0105] 3.3 Solution preparation

[0106] PBS buffer (pH 7.4): Dissolve PBS powder (1 L) in 1000 mL of purified water, mix well, sterilize with high-pressure steam at 121°C for 30 min, and store at 4°C for later use.

[0107] Cell digestion solution: Dissolve 0.25% Trypsin and 0.02% EDTA in PBS buffer (pH 7.4), pH 7.8-8.0, filter with a 0.22 μm filter, and store at 4°C for future use.

[0108] High-glucose DMEM medium: Dissolve high-glucose DMEM powder with ultrapure water, pH 7.2, filter with a 0.22 μm filter, add 10% FBS before use, and store at 4°C for later use.

[0109] 1.5mMH2O2: Take 42.5μL of 3% hydrogen peroxide and add it to 25mL of sterile PBS to prepare a stock solution with a concentration of 1.5mmol / L. Dilute it with culture medium to a concentration of 250μmol / L and use it immediately after preparation.

[0110] DCFH-DA: The working solution was prepared at a dilution ratio of 1:1000, and DCFH-DA was diluted with PBS to a final concentration of 10 μmol / L.

[0111] 4. Instruments and Equipment

[0112] Electronic balance (Pioneer), vortex machine (Hangzhou Miou), pipette (Eppendorf), CO2 cell culture incubator (Thermo), cell counter (Invitrogen), inverted fluorescence microscope (Nikon), oscillator (Kylin-Bell), clean bench (Sujing Antai), low-temperature high-speed centrifuge (Eppendorf), flow cytometer (Beckman).

[0113] 5. Detection Methods

[0114] 5.1 Sample preparation

[0115] The target peptide stock solution with a concentration of 2 mg / mL was diluted 80 times and 160 times to prepare a sample solution with the required dilution multiple.

[0116] 5.2 Test steps

[0117] Reactive oxygen species (ROS) detection: HaCaT cells in the logarithmic growth phase were cultured at 4 × 10 5 The cells were inoculated into 6-well plates at a density of 100 / well and incubated overnight in an incubator (37°C, 5% CO2). Experimental groups: The experiment was divided into a bare cell group, a blank group, a model group, and a sample group. Fresh culture medium was added to the bare cell group, the blank group (BC), and the model group (NC), and the sample group: the sample concentration was added.

[0118] H2O2-induced stimulation: Wash twice, add 250μM H2O2 to the model group and sample group for stimulation, 2mL per well, incubate in an incubator (37°C, 5% CO2) for 30min, add 10% FBS high-glucose medium to the bare cell group and blank group. After the stimulation, add 2mL / well PBS for washing twice.

[0119] Loading probe: Except for the bare cell group, add 1 mL of 10 μmol / L DCFH-DA to each well. Incubate in a 37°C cell culture incubator for 30 min. After the DCFH-DA incubation is complete, add 1 mL of PBS solution to each well to gently wash the cells, repeat 3 times.

[0120] Cell digestion: Add 700 μL of trypsin to each well and digest the cells at 37°C for 3 minutes. Stop digestion and add 700 μL of DMEM culture medium containing 10% newborn calf serum to each well. Blow repeatedly to ensure that all cells are blown down. Transfer the cell suspension after centrifugation into a 1.5mL Eppendorf tube, centrifuge at 3000r / min for 3 minutes, and discard the supernatant. Add 1mL PBS to each tube to wash the cells once and discard the supernatant. Add 500 μL PBS to each tube to resuspend the cells and detect by flow cytometry.

[0121] 5.4 Statistics

[0122] The software was used to perform a T-Test on the values ​​of the sample group and the model group. P < 0.05 was considered to be a significant statistical difference, indicated by *, and P < 0.01 was considered to be an extremely significant statistical difference, indicated by **.

[0123] 6Test results

[0124] As shown in Table 3 and Figure 5 As shown, the sample acted on HaCaT cells for 24 hours. Compared with the model group, the ROS inhibition rate of the target peptide diluted 160 times and 80 times in the sample group was significantly reduced, with inhibition rates of 25.39% and 57.09%. The ROS content in the model group was increased by ≥30% compared with the blank group / model group, and this experiment met the standards. To evaluate the ability of the test substance to inhibit ROS, the inhibition rate value of the test substance needs to be positive, indicating that the sample test substance has the effect of inhibiting ROS at this concentration, and also indicates that it has a firming and anti-wrinkle effect. And the green fluorescence intensity (MFI) is proportional to the level of reactive oxygen species (ROS) in the cell.

[0125] Table 3 ROS inhibition rate detection results

[0126]

[0127] “ / ” means the item is not included.

[0128] It should be noted that when the claims of the present invention involve numerical ranges, it should be understood that the two endpoints of each numerical range and any numerical value between the two endpoints can be selected. In order to avoid redundancy, the present invention describes a preferred embodiment.

[0129] Although the preferred embodiments of the present invention have been described, those skilled in the art may make other changes and modifications to these embodiments once they have learned the basic creative concept. Therefore, the appended claims are intended to be interpreted as including the preferred embodiments and all changes and modifications that fall within the scope of the present invention.

[0130] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. A polypeptide that can remove reactive oxygen species and increase SIRT gene expression, characterized in that: The sequence of the polypeptide is nicotinic acid-CCArBrBrBrARGDA-NH2; wherein G is glycine, R and r are arginine, D is aspartic acid, A is tyrosine, B is tryptophan, and C is phenylalanine. In the polypeptide, r is in D configuration, and the remaining amino acids are in L configuration.

2. Use of the polypeptide according to claim 1 in the preparation of cosmetics or medicines for resisting the aging of human skin cells.

3. A drug containing the polypeptide according to claim 1, characterized in that: The polypeptide exists in the form of acetate.

4. The drug according to claim 3, characterized in that The medicine also contains pharmaceutically acceptable excipients.

5. The drug according to claim 4, characterized in that The auxiliary material is an auxiliary material required for preparing an aqueous extract, a powder, a lotion, a tincture, an oil, an emulsion, an ointment, a plaster or an aerosol.

6. A cosmetic containing the polypeptide according to claim 1.

7. The cosmetic according to claim 6, characterized in that: The cosmetic is one of essence, toner, lotion or cream.

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