Polypeptide, Anti-aging transdermal hydrogel, and use

Abstract

Provided are a polypeptide, an anti-aging transdermal hydrogel, and a use, relating to the fields of biomedicine and novel drug development. The sequence of the polypeptide is (C12)-FGGRGGHGGGG-(3-Aminobenzeneboronic acid), wherein C12 represents lauric acid, 3-Aminobenzeneboronic acid represents 3-aminophenylboronic acid, F represents phenylalanine, G represents glycine, R represents arginine, and H represents histidine. The polypeptide can be chemically modified and self-assembled in an aqueous solution to form a supramolecular hydrogel. Experiments show that the polypeptide hydrogel has strong transdermal permeability, can reach the corium layer within2 h, can significantly inhibit aging phenotypes such as transcription of SASP factors, activation of P53, up-regulation of β-Gal, and expression of γH2AX, can be used for preparing anti-aging products, and has good market prospects and application potential.

Description

A polypeptide, a transdermal hydrogel for anti-aging and its applications Technical Field

[0001] This invention relates to the fields of biomedicine and novel drug development, specifically to a polypeptide, a transdermal hydrogel for anti-aging, and their applications. Background Technology

[0002] Human skin aging is caused by natural or unnatural factors. The growth period of skin tissue generally ends around the age of 25. After that, growth and aging occur simultaneously. The elastic fibers of the skin gradually thicken, and as age continues to increase, skin aging becomes more and more obvious. Skin aging is usually reflected in several aspects, such as skin elasticity, wrinkles and roughness, and the degree of moisturization. In addition, environmental factors, sunlight, and stress can cause free radicals to damage the skin, accelerating skin aging and making the skin more prone to spots, fine lines, and other signs of aging.

[0003] Anti-aging peptides are widely studied compounds that can treat skin aging by promoting collagen production, acting as antioxidants, and inhibiting protease activity, making them ideal anti-aging drugs. However, due to the skin's natural protective barrier function and limited permeability in transporting pathogens and chemicals from the external environment, the penetration of anti-aging peptides across the skin barrier has always been a major challenge to overcome. Furthermore, anti-aging peptides may lose activity during transdermal delivery due to environmental factors (such as oxidative degradation); therefore, enhancing and protecting their stability is crucial to ensuring the effectiveness of peptides during delivery.

[0004] Therefore, there is an urgent need in this field to develop an anti-aging product with strong transdermal penetration, significant anti-aging effects, and stable efficacy. Summary of the Invention

[0005] This invention addresses the problem of the lack of anti-aging products with strong transdermal penetration, significant and stable anti-aging effects in the existing technology. It provides a polypeptide, a transdermal hydrogel for anti-aging, and its application. The hydrogel prepared using this polypeptide not only has strong transdermal penetration, reaching the dermis layer in 2 hours, but also significantly inhibits aging phenotypes such as SASP factor transcription, p53 activation, β-Gal upregulation, and γH2AX expression. Therefore, it can be used to prepare anti-aging products and has good market prospects and application potential.

[0006] Based on the above, the present invention first provides a polypeptide, the structure of which is shown below:

[0007] In another aspect, the present invention provides a transdermal hydrogel for anti-aging, the hydrogel comprising the aforementioned polypeptide.

[0008] In another aspect, the present invention provides a method for preparing the aforementioned transdermal hydrogel for anti-aging, comprising the following steps:

[0009] S1, Weigh the aforementioned polypeptide and dissolve it in water. After mixing evenly, obtain an aqueous polypeptide solution.

[0010] S2, dissolve the polypeptide in an aqueous solution under ultrasonic conditions;

[0011] S3. The aqueous solution after step S2 is left to stand at room temperature for at least 10 minutes to obtain the transdermal hydrogel for anti-aging.

[0012] Preferably, the concentration of the polypeptide in the aqueous solution is 5 mg / mL to 15 mg / mL.

[0013] Preferably, the ultrasonic conditions are: ultrasonic power 80w-150w, ultrasonic time 20s-40s, and ultrasonic temperature 20℃-35℃.

[0014] Preferably, in step S1, the polypeptide weighed is a lyophilized polypeptide powder, and the purity of the lyophilized polypeptide powder is ≥95%.

[0015] Preferably, in step S1, the water used is deionized water.

[0016] The aforementioned peptides or hydrogels are used in the preparation of anti-aging products.

[0017] Preferably, the anti-aging is anti-skin aging.

[0018] Preferably, the method of using the anti-aging product includes transdermal administration.

[0019] Compared with the prior art, the beneficial effects of the present invention include at least the following:

[0020] This invention provides a polypeptide that can self-assemble into a supramolecular hydrogel in aqueous solution through chemical modification. This allows for effective delivery of the polypeptide to the surface or deeper layers of the skin, enhancing its stability and activity. A series of cell and animal model experiments evaluated the transdermal and anti-aging effects of this polypeptide hydrogel: the hydrogel exhibits strong transdermal penetration, reaching the dermis within 2 hours, and significantly inhibits aging phenotypes such as SASP factor transcription, p53 activation, β-Gal upregulation, and γH2AX expression. It can be used to prepare anti-aging products and has promising market prospects and application potential. Attached Figure Description

[0021] Figure 1 is a photograph of an inverted / tilted polypeptide hydrogel according to an embodiment of the present invention, wherein:

[0022] A is an inverted photograph of the polypeptide aqueous solution of the present invention after it has been allowed to stand.

[0023] B is a tilted photograph of the control polypeptide aqueous solution after it has been left to stand.

[0024] Figure 2 is a transmission electron microscope image of a polypeptide hydrogel according to an embodiment of the present invention.

[0025] Figure 3 shows the infrared spectra of a polypeptide hydrogel before and after gelation according to an embodiment of the present invention.

[0026] Figure 4 shows the cytotoxicity experiment of the polypeptide hydrogel of the present invention.

[0027] Figure 5 is an immunoblot image of the peptide hydrogel of the present invention inhibiting the aging-related gene P53.

[0028] Figure 6 shows the expression of galactosidase inhibition by the polypeptide hydrogel of the present invention.

[0029] Figure 7 is a confocal image of the peptide hydrogel of the present invention inhibiting the expression of DNA damage γH2AX.

[0030] Figure 8 shows the quantitative expression of γH2AX in the polypeptide hydrogel of the present invention.

[0031] Figure 9 shows the PCR fluorescence quantitative PCR spectrum of the peptide hydrogel of the present invention for inhibiting aging phenotype-related factors, wherein:

[0032] A is the Tnf-α PCR fluorescence quantitative chromatogram;

[0033] B is the quantitative PCR chromatogram of Cxcl10.

[0034] Figure 10 shows the transdermal effect of the polypeptide hydrogel of the present invention after 2 hours of application.

[0035] Figure 11 shows the anti-aging effect of the polypeptide hydrogel of the present invention. Detailed Implementation

[0036] The technical solution of the present invention will be further described in detail below with reference to the accompanying drawings and embodiments.

[0037] As mentioned earlier, while peptides are ideal anti-aging drugs, their poor transdermal delivery and stability pose a significant challenge. To address this issue, this invention, through extensive research and experimentation, ultimately combines anti-aging peptides with hydrogel technology. Firstly, it provides a peptide with anti-aging effects. Secondly, this peptide can self-assemble into a supramolecular hydrogel in aqueous solution through chemical modification. The hydrogel, acting as an excellent carrier, can effectively deliver the anti-aging peptide to the skin surface or deeper layers, improving its stability and activity, thereby enhancing the anti-aging therapeutic effect.

[0038] Therefore, the first object of the present invention is to provide a polypeptide with the sequence: (C12)-FGGRGGHGGGG-(3-Aminobenzeneboronic acid), where C12 represents lauric acid, 3-Aminobenzeneboronic acid represents 3-aminophenylboronic acid, F represents phenylalanine, G represents glycine, R represents arginine, and H represents histidine. The structure of the polypeptide is shown below:

[0039] A second object of the present invention is to provide a transdermal hydrogel for anti-aging, the hydrogel comprising the aforementioned polypeptide.

[0040] A third objective of this invention is to provide a method for preparing the aforementioned transdermal hydrogel for anti-aging, comprising the following steps:

[0041] S1, Weigh the aforementioned polypeptide and dissolve it in water. After mixing evenly, obtain an aqueous polypeptide solution.

[0042] S2, Dissolve the polypeptide aqueous solution from step S1 under ultrasonic conditions;

[0043] S3. The aqueous solution after step S2 is left to stand at room temperature for at least 10 minutes to obtain the transdermal hydrogel for anti-aging.

[0044] In some embodiments, the concentration of the polypeptide in the aqueous solution is 5 mg / mL to 15 mg / mL.

[0045] In some embodiments, the ultrasonic conditions are: ultrasonic power 80w-150w, ultrasonic time 20s-40s, and ultrasonic temperature 20℃-35℃.

[0046] In some embodiments, in step S1, the polypeptide weighed is a polypeptide lyophilized powder with a purity ≥95%; and the water used is deionized water.

[0047] A fourth objective of this invention is to provide the application of the aforementioned polypeptide or the aforementioned hydrogel in the preparation of anti-aging products.

[0048] In some embodiments, the anti-aging is anti-skin aging.

[0049] In some embodiments, the method of using the anti-aging product includes transdermal delivery.

[0050] The research process and results of this invention will be described in detail below with reference to experimental data:

[0051] Unless otherwise stated, the experimental methods, detection methods, and preparation methods disclosed in this invention all employ conventional techniques in molecular biology, biochemistry, chromatin structure and analysis, analytical chemistry, cell culture, recombinant DNA technology, and related fields. Unless otherwise specified, the experimental methods used in the following examples were performed under conventional or manufacturer-recommended conditions.

[0052] Unless otherwise specified, all materials and reagents used in the following examples are commercially available.

[0053] (I) Polypeptide Synthesis

[0054] The following peptide structure was synthesized using solid-phase synthesis technology. The specific synthesis was completed by the synthesis service provider according to the provided structure, and it was delivered in lyophilized powder form.

[0055] Third-party structural testing confirmed the polypeptide sequence as: (C12)-FGGRGGHGGGG-(3-Aminobenzeneboronic acid), which is the structure shown below, and matches the target structure.

[0056] This embodiment also provides a comparative peptide for comparing hydrogel-forming abilities. The sequence of the comparative peptide is: FGGRGGHGGGG-(3-Aminobenzeneboronic acid), and the structure of the comparative peptide is shown below:

[0057] Analysis shows that, compared to the structure of the polypeptide of the present invention, the 5' end of the contrast polypeptide sequence is not modified with lauric acid.

[0058] (II) Preparation and Characterization of Hydrogels

[0059] 1. Preparation of hydrogels

[0060] Weigh 5 mg of the lyophilized peptide powder of the present invention or the comparative lyophilized peptide powder and mix it with 0.5 mL of deionized water; then dissolve it by sonication at room temperature with a power of 100 W and a duration of 30 s; after sonication is complete, let it stand for 10 minutes and observe whether a hydrogel has formed by inverting or tilting the vial.

[0061] The results are shown in Figure 1. The inverted vial demonstrates that the polypeptide aqueous solution of the present invention can form a non-flowing hydrogel state after dissolution and standing (Figure 1A); while the polypeptide aqueous solution vial continues to flow after being tilted, and it cannot form a hydrogel state (Figure 1B).

[0062] The above results indicate that the introduction of lauric acid plays an important role in the formation of hydrogels from the aqueous solution of the polypeptides of this invention, and is an important component of the polypeptide structure of this invention.

[0063] 2. Characterization of hydrogels

[0064] TEM morphology characterization: The polypeptide aqueous solution prepared according to the above method was shaken well, and 10 μL was dropped onto a 200-mesh carbon support film. After drying, it was stained with 1% phosphotungstic acid solution for about 30 seconds, and then dried for TEM testing. The results are shown in Figure 2, which shows that the polypeptide hydrogel of the present invention is formed by dense nanofibers intertwined with each other.

[0065] Infrared spectroscopy determination: The polypeptide hydrogel prepared by the above method was freeze-dried at -50℃ to obtain a lyophilized hydrogel powder. Potassium bromide was added and pressed into a pellet (the ratio of lyophilized powder to potassium bromide was 1:20), and the infrared spectrum was measured. The lyophilized polypeptide powder was included as a control. The results are shown in Figure 3. Compared with the infrared spectrum of the lyophilized polypeptide powder, the infrared absorption wavelength of the lyophilized hydrogel powder showed a red shift, indicating that intermolecular hydrogen bonding occurs in the gel state.

[0066] (III) Evaluation of the in vitro anti-aging effects of hydrogels

[0067] 1. Cytotoxicity detection

[0068] Human immortalized epidermal cells (HaCaT cells) were used to investigate the cytotoxicity of the hydrogel. Different concentrations (0 μg / mL, 5 μg / mL, 10 μg / mL, 15 μg / mL, 20 μg / mL) of peptide hydrogel solutions were prepared and sterilized under UV irradiation for 12 h before use. HaCaT cells were seeded at a density of 20,000 cells / well in 96-well plates and cultured in a CO2 incubator using 10% fetal bovine serum and antibiotic-containing DMEM as the complete growth medium. After 24 h of culture, the hydrogel was added to the wells. After 1, 3, and 5 days of culture, the cell culture medium was replaced with 100 μL of fresh DMEM. After 5 days of culture, 10% (v / v) CCK-8 reagent (Beyotime, China) was added simultaneously. After 2 h of culture at 37°C, cell viability was quantified using a 450 nm microplate reader (TECAN, Switzerland).

[0069] As shown in Figure 4, compared with the control group (0 μg / mL), there was no significant difference in cell viability among the experimental groups containing different concentrations of hydrogel, indicating that the hydrogel of the present invention has no obvious toxic effect on cells and has good biocompatibility.

[0070] 2. Evaluation of anti-aging effects

[0071] After multiple passages, the cells exhibit senescent cell phenotypes, such as increased expression of senescence-associated secretory phenotypic factors (SASPs) TNF-α and Cxcl10, high expression of β-galactosidase (β-Gal), nuclear DNA damage, and elevated expression of cyclin P53. By isolating mouse primary embryonic fibroblasts (MEFs) and detecting changes in the expression of relevant senescence factors or markers during cell senescence from P1 to P3-P4 passages, the anti-aging effect of the polypeptide hydrogel of this invention was evaluated.

[0072] The method for isolating mouse primary embryonic fibroblasts (MEFs) is as follows: Each pregnant mouse (13-14 days old) is injected intraperitoneally with 0.5 mL of avertin. After anesthesia, the mice are euthanized by cervical dislocation and disinfected by immersion in 75% ethanol for 2-5 minutes. The abdominal skin of the pregnant mouse is cut open with sterile scissors and forceps to expose the abdominal cavity. The uterus is removed and placed in a pre-prepared sterile culture dish. The carcass of the pregnant mouse is disposed of. The uterus is rinsed three times with 10 mL PBS using a pipette. The blastocyst is cut open with ophthalmic scissors and forceps to free the embryo. The embryo is placed in a new sterile culture dish and washed three times with 10 mL PBS. The number of isolated embryos is counted. The head, tail, limbs, and internal organs of the embryo are removed. The embryo is placed in a new culture dish and washed with 10 mL PBS. Wash three times with PBS; remove excess PBS, and use curved ophthalmic scissors to cut the tissue into small particles, taking about 6 to 10 minutes to cut the tissue; add 2 mL of trypsin and continue cutting for a few minutes to make the tissue particles even smaller; add another 5 mL of trypsin, and use a pipette to mix the tissue and trypsin thoroughly, and incubate the culture dish at 37°C for 20 to 30 minutes, during which time you can take it out and repeatedly blow and aspirate it several times with a sterile 25 mL pipette. Note: During incubation, another mouse can be processed and prepared starting from step one; after digestion, add approximately 20 mL of MEF-I medium, mix the viscous substance with a 10 mL pipette to disperse it, and aspirate the cell suspension. Do not digest the remaining clumps with trypsin, otherwise cell viability will decrease; filter through a 200-mesh sieve, being careful not to let the viscous substance clog the mesh; after filtration, transfer the tissue digestion solution to a 50 mL centrifuge tube and centrifuge at 200 g for 5 min; discard the supernatant, resuspend in MEF-C medium, and aliquot into T75 culture flasks for primary culture, with each flask containing approximately 10 cells obtained from embryo digestion; culture overnight and observe under a microscope. If cell confluence exceeds 90%, cells can be directly collected and cryopreserved.

[0073] The experimental process and results for detecting the anti-aging effect of the polypeptide hydrogel of the present invention are as follows:

[0074] (1) Inhibition of P53 activation: Mouse primary embryonic fibroblasts (MEFs) were isolated and passaged continuously to obtain P1-P3 generation MEF cells. These cells were cultured in 24-well plates for 24 hours, and then the polypeptide hydrogel of this invention was added (gel1 group: 10 μL of the prepared polypeptide hydrogel was added; Ctrl group: the same volume of water was added). When the cells reached ideal confluence, the culture medium was aspirated, and the cells were washed three times with PBS to remove excess medium. Then, 100 μL of RAPI lysis buffer and protease inhibitor were added to extract total protein, which was quantified using a BCA protein assay kit. 30 μg of protein sample was dissolved in 10% SDS-PAGE and transferred to a PVDF membrane. The membrane was incubated overnight at 4°C with P53 primary antibody (1:500, Abcam), using β-actin as an internal control protein. The PVDF membrane was then immersed in 5% skim milk for 1 hour and incubated with secondary antibody at room temperature for 2 hours. Finally, the membrane was washed three times with TBST buffer, and the blot was observed using an enhanced chemiluminescence detection system.

[0075] As shown in Figure 5, with passage, the P53 protein content of the P3 generation cells in the gel1 group was significantly reduced compared to the P3 generation cells in the Ctrl group, indicating that the polypeptide hydrogel of the present invention can inhibit the activation of the P53 gene.

[0076] (2) Inhibition of β-galactosidase expression: Primary mouse embryonic fibroblasts (MEFs) were isolated and passaged continuously to obtain P4 generation MEF cells. Cells were seeded in 12-well plates with / without the polypeptide hydrogel of this invention (Gel1 group: 10 μL of the prepared polypeptide hydrogel was added; H2O group: the same volume of water was added). After culture, the β-galactosidase (SA-β-Gal) content was measured using a β-galactosidase kit. (It should be noted that SA-β-Gal, β-Gal, and β-galactosidase in this invention are the same substance and can be used interchangeably.)

[0077] As shown in Figure 6, with passage, the content of SA-β-Gal protein in the P4 generation cells of the Gel1 group was significantly reduced compared with that of the H2O group, indicating that the polypeptide hydrogel of the present invention can reduce the expression of β-galactosidase.

[0078] (3) Inhibition of DNA damage: Mouse primary embryonic fibroblasts (MEFs) were isolated and passaged continuously to obtain P4 generation MEF cells. The cells were seeded in confocal culture dishes with / without the polypeptide hydrogel of this invention (gel1 group: 10 μL of the polypeptide hydrogel prepared above was added; Ctrl group: the same volume of water was added). After culture, the expression of the DNA damage marker γH2AX was measured using a DNA damage kit.

[0079] As shown in Figures 7 and 8, compared with the Ctrl group, the expression level of γH2AX in the gel1 group cells was significantly reduced, indicating that the polypeptide hydrogel of the present invention can significantly inhibit DNA damage and delay cell senescence.

[0080] (4) Inhibit the transcription of aging-related secretory phenotypic factors (SASPs): 1×10 6 Primary embryonic fibroblast (MEF) cells from generations P1-P4 of mice were seeded in 12-well plates and cultured for 24 h. Then, 10 μL of the polypeptide hydrogel prepared according to this invention (Gel1 group) or the same volume of water (H2O group) was added to the wells, and the cells were cultured at 37°C for 2 days. Total RNA was extracted using the Ezscript reverse transcription kit (EZ Bioscience, USA) according to the manufacturer's instructions, and then reverse transcribed into cDNA using the SYBR PrimerScript RT-PCR kit. The synthesized cDNA was stored at -80°C. The mRNA levels of TNF-α and Clcx10 in each sample were detected by real-time quantitative PCR using the following method: 95°C for 5 min, 1 cycle; 95°C for 10 s; 60°C for 30 s, 40 cycles. The relative mRNA levels of each gene were calculated using the 2-ΔΔCt method with β-actin as the reference gene. Each reaction was performed three times.

[0081] As shown in Figure 9, with passage, the levels of TNF-α and Cxcl10 mRNA in the P4 generation cells of the Gel1 group were significantly reduced compared to those in the H2O group.

[0082] The above results indicate that the polypeptide hydrogel of the present invention can significantly inhibit aging phenotypes such as transcription of SASP factor, activation of P53, upregulation of β-Gal, and expression of γH2AX in cells, and has a significant delaying effect on cell senescence.

[0083] (iv) Evaluation of the in vivo transdermal and anti-aging effects of hydrogels

[0084] 1. Transdermal test

[0085] Several 6-8 week old C57 mice were anesthetized, and their back hair was removed. A 1cm×2cm area was taken and 25μL of peptide hydrogel mixed with Cy5 fluorescence (10mg / mL) was applied. Samples were taken and embedded in OCT after 5, 20, 60 and 120 min of application. After being transferred to -80℃ overnight, the samples were ice-cut, mounted and photographed using laser confocal microscopy. The absorption of peptide hydrogel by mice was compared by fluorescence intensity.

[0086] As shown in Figure 10, compared with the control group, the fluorescence of the subcutaneous tissue of mice in the polypeptide hydrogel group (C12 group) was significantly enhanced, indicating that the subcutaneous tissue absorbed more polypeptide hydrogel, and the transdermal effect of the polypeptide hydrogel of the present invention was obvious.

[0087] 2. Evaluation of in vivo anti-aging effects

[0088] Fifteen nude mice (balb / c, 4-6 weeks old) were ordered and divided into three groups: a blank control group (Blank group), a PBS group, and a peptide hydrogel group (C12), with five mice in each group. The PBS and peptide hydrogel groups used UVB (outdoor ultraviolet light) and subcutaneous injection of galactose to induce a mouse aging model: the nude mice were anesthetized daily and injected subcutaneously with galactose on their backs, followed by 1 hour of UVB light exposure until aging skin conditions appeared. In the peptide hydrogel group, after daily light exposure and galactose injection, peptide hydrogel was also applied to the backs of the mice. The blank control group received no treatment.

[0089] As shown in Figure 11, on the fourth day, the skin on the backs of nude mice in the PBS group showed obvious signs of aging, while the skin of nude mice in the C12 group, which was treated with polypeptide hydrogel, did not show obvious signs of aging and was the same as the skin of the untreated control group. This indicates that the polypeptide hydrogel of the present invention has a significant anti-aging effect in anti-aging model mice.

[0090] In summary, this invention provides a polypeptide that can self-assemble into a supramolecular hydrogel in aqueous solution through chemical modification. A series of cell and animal model experiments were conducted to evaluate the transdermal and anti-aging effects of this polypeptide hydrogel. The polypeptide hydrogel of this invention exhibits strong transdermal penetration, reaching the dermis within 2 hours, and significantly inhibits aging phenotypes such as SASP factor transcription, p53 activation, β-Gal upregulation, and γH2AX expression. It can be used to prepare anti-aging products and has broad market prospects and application potential.

[0091] Although the present invention has been described in detail through the preferred embodiments above, it should be understood that the above description should not be considered as a limitation of the present invention. Various modifications and substitutions to the present invention will be apparent to those skilled in the art after reading the above description. Therefore, the scope of protection of the present invention should be defined by the appended claims.

Claims

1. A polypeptide, characterized in that, The structure of the polypeptide is as shown below:

2. A transdermal hydrogel for anti-aging, characterized by, The hydrogel comprises the polypeptide of claim 1.

3. The method for the preparation of transdermal hydrogel for anti-aging as claimed in claim 2, wherein, The method comprises the following steps: S1, the polypeptide of claim 1 is weighed and dissolved in water, and after uniform mixing, a polypeptide aqueous solution is obtained; S2, the polypeptide aqueous solution is dissolved under ultrasonic conditions; S3, the aqueous solution after step S2 is treated is left to stand at room temperature for at least 10 minutes to obtain the transdermal hydrogel for anti-aging.

4. The production method according to claim 3, wherein The concentration of the polypeptide in the polypeptide aqueous solution is 5 mg / mL-15 mg / mL.

5. The production method according to claim 3, wherein The ultrasonic conditions are: ultrasonic power 80 w-150 w, ultrasonic time 20 s-40 s, and ultrasonic temperature 20℃-35℃.

6. The production method according to claim 3, wherein In step S1, the weighed polypeptide is a polypeptide freeze-dried powder, and the purity of the polypeptide freeze-dried powder is ≥95%.

7. The production method according to claim 3, wherein In step S1, the water used is deionized water.

8. Use of the polypeptide of claim 1 or the hydrogel of claim 2 in the preparation of an anti-aging product.

9. Use according to claim 8, wherein the compound is ###0002### The anti-aging is anti-skin aging.

10. The use according to claim 8, wherein the compound is ###00002### The use method of the anti-aging product comprises transdermal administration.

Images