Stapled peptide compound, preparation method and application thereof, and skin care product

Abstract

The invention relates to the technical field of polypeptide compounds, in particular to a stapled peptide compound, a preparation method and application thereof and a skin care product. The structural formula of the stapled peptide compound is shown in the specification. The polypeptide has high anti-hydrolysis stability, can significantly improve the expression ability of up-regulated fiber cells and promote the expression of collagen, and can be used in the fields of wrinkle resistance, aging resistance, repair and the like.

Description

Technical Field

[0001] This invention relates to the field of peptide compound technology, and more specifically, to peptide compounds, their preparation methods, their applications, and skincare products. Background Technology

[0002] Peptide compounds are organic compounds formed by the dehydration condensation of multiple amino acids through peptide bonds (amide bonds). They possess advantages such as high target affinity and specificity, low toxicity and side effects, weaker immunogenicity than antibodies, and superior tissue permeability compared to macromolecules, leading to their widespread application in anti-aging and skin repair. However, traditional peptide compounds are easily degraded under physiological conditions, which affects the long-term efficacy of their drugs.

[0003] In view of this, the present invention is proposed. Summary of the Invention

[0004] The purpose of this invention is to provide a staple peptide compound, its preparation method, its application, and skincare products. This invention provides a novel staple peptide compound with high hydrolytic stability, which can significantly enhance the upregulation of fibroblast expression and promote collagen expression, and can be used in anti-wrinkle, anti-aging, and repair applications.

[0005] This invention is implemented as follows: In a first aspect, the present invention provides a staple peptide compound having the following structural formula: .

[0006] Secondly, the present invention provides a method for preparing the staple peptide compound described in the foregoing embodiments, comprising: staplering two side chain residues of a linear polypeptide to form the staple peptide compound; wherein the amino acid sequence of the linear polypeptide is as follows: AC-Lys-Glu-Pro-Pro-Pro-Lys-Arg-Pro-Ala-Asp-NH2.

[0007] In an optional embodiment, the binding process includes: reacting the linear polypeptide with activated glutaric acid in a catalyst system.

[0008] In an optional embodiment, the glutaric acid is activated by means of any of the following substances: bis(succinimide), pentafluorophenyl ester, dicyclohexylcarbodiimide carbonate, hydroxybenzotriazole, and azide.

[0009] In an optional embodiment, the molar ratio of the linear polypeptide to the activated glutaric acid is 1:(1-1.5).

[0010] In an optional embodiment, the catalyst includes at least one of DIC, HOBt, and DIEA.

[0011] In an optional embodiment, the molar ratio of the linear polypeptide to the catalyst is 1:(5-10).

[0012] Thirdly, the present invention provides the use of the staple peptide compound described in the foregoing embodiments in the preparation of a promoter that promotes collagen expression.

[0013] Fourthly, the present invention provides the application of the staple peptide compound described in the foregoing embodiments in the preparation of anti-wrinkle, anti-aging and / or repairing skin care products.

[0014] Fifthly, the present invention provides a skin care product comprising the staple peptide compound described in the foregoing embodiments.

[0015] The present invention has the following beneficial effects: The embodiments of the present invention provide a staple peptide compound with high stability, particularly high resistance to hydrolysis, thereby ensuring the long-term effectiveness of the staple peptide compound. Simultaneously, this staple peptide compound can significantly upregulate fibroblast expression and enhance the expression of various collagen proteins, thus enabling its use in anti-wrinkle, anti-aging, and repair applications. Attached Figure Description

[0016] 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.

[0017] Figure 1 This is a high-performance liquid chromatography (HPLC) spectrum of the staple peptide compound provided in Example 1 of the present invention; Figure 2 The mass spectrum of the staple peptide compound provided in Example 1 of this invention; Figure 3 The test result diagram provided for Test Example 1 of the present invention; Figure 4 The test result diagram provided for Test Example 2 of the present invention; Figures 5-12 The test result diagram provided for Test Example 3 of the present invention. Detailed Implementation

[0018] 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.

[0019] In a first aspect, the present invention provides a staple peptide compound having the following structural formula: , The amino acid sequence of this staple peptide compound is as follows: AC-Lys-Glu-Pro-Pro-Pro-Lys-Arg-Pro-Ala-Asp-NH2 (Lys2 & Lys7 by Glutaric Acid). This staple peptide compound exhibits high resistance to hydrolysis; for example, it is not easily degraded under physiological conditions, thus prolonging its duration of action, i.e., it has a long physiological half-life. Simultaneously, this staple peptide compound can significantly upregulate fibroblast expression and enhance the expression of various collagen proteins, potentially providing a more efficient and safer option for the cosmetic and medical fields.

[0020] Secondly, the present invention provides a method for preparing the staple peptide compound described in the foregoing embodiments, comprising: stapled two side chain residues of a linear polypeptide to form the staple peptide compound; wherein the amino acid sequence of the linear polypeptide is as follows: AC-Lys-Glu-Pro-Pro-Pro-Lys-Arg-Pro-Ala-Asp-NH2. This linear polypeptide can be prepared in-house or can be purchased directly from existing commercial products.

[0021] Linear polypeptides can be prepared in-house using the following methods: A fully protected peptide resin containing linear polypeptides in its sequence was prepared using conventional synthetic methods in the field of peptides. These conventional synthetic methods include solid-phase synthesis and liquid-phase synthesis.

[0022] The steps of the solid-phase synthesis method provided in this embodiment of the invention include: Coupling of protected amino groups: The first protecting amino group is covalently coupled to the solid support. Deprotection: Removing the protecting group on the amino group under stable conditions to expose the free amino group. Coupling: The process of condensing the next activated amino acid with the previous amino acid bound to the solid phase to form a peptide bond. Peptide bond elongation: Repeatedly perform two steps of deprotection and coupling to elongate the peptide chain to the designed length. Cleavage: The synthesized peptide is cleaved from the solid support and the side chain protection is removed.

[0023] It should be noted that the solid-phase synthesis provided in the embodiments of the present invention is only a general outline of the steps. The above steps may be added or reduced as appropriate. In addition, the specific operating conditions are all conventional operations in the art and will not be described in detail in the embodiments of the present invention.

[0024] Similarly, the liquid-phase synthesis steps provided in the embodiments of the present invention include: Amino acid protection: Protecting the functional groups of amino acids to prevent side reactions during the reaction process. Activation: An activator is used to couple the protected amino acids with the amino acids from the previous step to form peptide bonds. Coupling: Amino acids are linked together using a linker (such as DCC) to form a peptide chain. Deprotection: Removing the protecting group to elongate the peptide chain. Repeat: These steps need to be repeated until the target polypeptide compound is synthesized.

[0025] It should be noted that the liquid-phase synthesis provided in the embodiments of the present invention is only a general outline of the steps. The above steps may be added or reduced as needed. In addition, the specific operating conditions are all conventional operations in the art and will not be described in detail in the embodiments of the present invention.

[0026] Furthermore, the linear polypeptide fully protected peptide resin is deprotected by cleavage to remove the protecting groups on the linear polypeptide fully protected peptide resin, resulting in a chain-like deprotected polypeptide, which is the linear polypeptide provided in the embodiments of the present invention.

[0027] Furthermore, the linear polypeptide is added to a solvent and reacted with activated glutaric acid in a catalyst system.

[0028] The following substances are used to activate glutaric acid: bis(succinimide), pentafluorophenyl ester, dicyclohexylcarbodiimide carbonate, hydroxybenzotriazole, and azide. The molar ratio of the linear polypeptide to the activated glutaric acid is 1:(1-1.5). For example, it can be any value between 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5, or 1:(1-1.5).

[0029] The catalyst includes at least one of DIC, HOBt, and DIEA. The molar ratio of the linear polypeptide to the catalyst is 1:(5-10). For example, it is any value between 1:5, 1:7, 1:10, or 1:(5-10).

[0030] Thirdly, the present invention provides a skincare product comprising the stapling peptide compound described in the foregoing embodiments. This skincare product has anti-wrinkle, anti-aging, and repairing effects. Furthermore, this skincare product includes, but is not limited to, toners, lotions, essences, lotions, creams, masks, sprays, and eye creams.

[0031] The present invention also provides the above-mentioned staple peptide compound with the following applications: (1) a promoter for promoting collagen expression; (2) a skin care product for anti-wrinkle, anti-aging and / or repair.

[0032] The features and performance of the present invention will be further described in detail below with reference to embodiments.

[0033] Example 1 This invention provides a method for preparing a staple peptide compound, comprising: S1: Swelling and Deprotection: Add 300 mmol of resin to the reactor, then add 1000 mL of 20% Piperidine / DMF solution. React at 25.0-28.0℃ for 30 min, and remove the liquid by filtration. Then wash 6 times with 1 L of DMF solution and dry for later use.

[0034] S2: Weigh Fmoc-Linker (485.6 g, 900 mmol) and HOBT (121.6 g, 900 mmol) into a dedicated activation vessel, dissolve in 1500 ml of DMF, maintain the temperature at 2-8℃, then slowly add DIC (139.4 ml, 900 mmol), activate for 5 min, then transfer to a reactor, turn on nitrogen, maintain the temperature at 25-28℃ for 45 min until the reaction is complete, filter to remove the liquid. Wash the resin three times with DMF solution, 1 L each time. After washing, add 1 L of 20% Piperidine / DMF solution, react at 25.0-28.0℃ for 30 min, filter to remove the liquid. Then wash six times with 1 L of DMF solution, dry and set aside for later use.

[0035] S3: Weigh 370.3 g (900 mmol) of Fmoc-Asp(OtBu)-OH and HOBT (121.6 g, 900 mmol) into a dedicated activation vessel, dissolve in 1500 ml of DMF, maintain the temperature at 2-8℃, then slowly add DIC (139.4 ml, 900 mmol), activate for 5 min, transfer to a reactor, turn on nitrogen, and react at 25-28℃ for 45 min until the reaction is complete. Filter to remove the liquid. Wash the resin three times with 1 L of DMF solution each time. After washing, add 1 L of 20% Piperidine / DMF solution, react at 25.0-28.0℃ for 30 min, and filter to remove the liquid. Then wash six times with 1 L of DMF solution and dry for later use.

[0036] S4: Weigh Fmoc-Ala-OH (280.2 g, 900 mmol) and HOBT (121.6 g, 900 mmol) into a dedicated activation vessel, dissolve in 1500 ml of DMF, maintain the temperature at 2-8℃, then slowly add DIC (139.4 ml, 900 mmol), activate for 5 min, transfer to a reactor, turn on nitrogen, maintain the temperature at 25-28℃ for 45 min until the reaction is complete, filter to remove the liquid. Wash the resin three times with DMF solution, 1 L each time. After washing, add 1 L of 20% Piperidine / DMF solution, react at 25.0-28.0℃ for 30 min, filter to remove the liquid. Then wash six times with 1 L of DMF solution, dry and set aside for later use.

[0037] S5: Weigh Fmoc-Pro-OH (303.6 g, 900 mmol) and HOBT (121.6 g, 900 mmol) into a dedicated activation vessel, dissolve in 1500 ml of DMF, maintain the temperature at 2-8℃, then slowly add DIC (139.4 ml, 900 mmol), activate for 5 min, transfer to a reactor, turn on nitrogen, maintain the temperature at 25-28℃ for 45 min until the reaction is complete, filter to remove the liquid. Wash the resin three times with DMF solution, 1 L each time. After washing, add 1 L of 20% Piperidine / DMF solution, react at 25.0-28.0℃ for 30 min, filter to remove the liquid. Then wash six times with 1 L of DMF solution, dry and set aside for later use.

[0038] S6: Weigh 583.9 g (900 mmol) of Fmoc-Arg(Pbf)-OH and HOBT (121.6 g, 900 mmol) into a dedicated activation vessel, dissolve in 1500 ml of DMF, maintain the temperature at 2-8℃, then slowly add DIC (139.4 ml, 900 mmol), activate for 5 min, transfer to a reactor, turn on nitrogen, maintain the temperature at 25-28℃ and react for 45 min until the reaction is complete, filter to remove the liquid. Wash the resin three times with 1 L of DMF solution each time. After washing, add 1 L of 20% Piperidine / DMF solution, react at 25.0-28.0℃ for 30 min, filter to remove the liquid. Then wash six times with 1 L of DMF solution, dry and set aside for later use.

[0039] S7: Weigh 421.7 g (900 mmol) of Fmoc-Lys(Boc)-OH and HOBT (121.6 g, 900 mmol) into a dedicated activation vessel, dissolve in 1500 ml of DMF, maintain the temperature at 2-8℃, then slowly add DIC (139.4 ml, 900 mmol), activate for 5 min, transfer to a reactor, turn on nitrogen, maintain the temperature at 25-28℃ for 45 min until the reaction is complete, and filter to remove the liquid. Wash the resin three times with 1 L of DMF solution each time. After washing, add 1 L of 20% Piperidine / DMF solution, react at 25.0-28.0℃ for 30 min, and filter to remove the liquid. Then wash six times with 1 L of DMF solution, and dry for later use.

[0040] S8: Weigh Fmoc-Pro-OH (303.6 g, 900 mmol) and HOBT (121.6 g, 900 mmol) into a dedicated activation vessel, dissolve in 1500 ml of DMF, maintain the temperature at 2-8℃, then slowly add DIC (139.4 ml, 900 mmol), activate for 5 min, transfer to a reactor, turn on nitrogen, maintain the temperature at 25-28℃ for 45 min until the reaction is complete, filter to remove the liquid. Wash the resin three times with DMF solution, 1 L each time. After washing, add 1 L of 20% Piperidine / DMF solution, react at 25.0-28.0℃ for 30 min, filter to remove the liquid. Then wash six times with 1 L of DMF solution, dry and set aside for later use.

[0041] S9: Weigh Fmoc-Pro-OH (303.6 g, 900 mmol) and HOBT (121.6 g, 900 mmol) into a dedicated activation vessel, dissolve in 1500 ml of DMF, maintain the temperature at 2-8℃, then slowly add DIC (139.4 ml, 900 mmol), activate for 5 min, transfer to a reactor, turn on nitrogen, maintain the temperature at 25-28℃ for 45 min until the reaction is complete, filter to remove the liquid. Wash the resin three times with DMF solution, 1 L each time. After washing, add 1 L of 20% Piperidine / DMF solution, react at 25.0-28.0℃ for 30 min, filter to remove the liquid. Then wash six times with 1 L of DMF solution, dry and set aside for later use.

[0042] S10: Weigh Fmoc-Pro-OH (303.6 g, 900 mmol) and HOBT (121.6 g, 900 mmol) into a dedicated activation vessel, dissolve in 1500 ml of DMF, maintain the temperature at 2-8℃, then slowly add DIC (139.4 ml, 900 mmol), activate for 5 min, transfer to a reactor, turn on nitrogen, maintain the temperature at 25-28℃ for 45 min until the reaction is complete, filter to remove the liquid. Wash the resin three times with DMF solution, 1 L each time. After washing, add 1 L of 20% Piperidine / DMF solution, react at 25.0-28.0℃ for 30 min, filter to remove the liquid. Then wash six times with 1 L of DMF solution, dry and set aside for later use.

[0043] S11: Weigh Fmoc-Glu(OtBu)-OH (382.9 g, 900 mmol) and HOBT (121.6 g, 900 mmol) into a dedicated activation vessel, dissolve in 1500 ml of DMF, maintain the temperature at 2-8℃, then slowly add DIC (139.4 ml, 900 mmol), activate for 5 min, transfer to a reactor, turn on nitrogen, maintain the temperature at 25-28℃ for 45 min until the reaction is complete, filter to remove the liquid. Wash the resin three times with DMF solution, 1 L each time. After washing, add 1 L of 20% Piperidine / DMF solution, react at 25.0-28.0℃ for 30 min, filter to remove the liquid. Then wash six times with 1 L of DMF solution, dry and set aside for later use.

[0044] S12: Weigh 421.7 g (900 mmol) of Fmoc-Lys(Boc)-OH and HOBT (121.6 g, 900 mmol) into a dedicated activation vessel, dissolve in 1500 ml of DMF, maintain the temperature at 2-8℃, then slowly add DIC (139.4 ml, 900 mmol), activate for 5 min, transfer to a reactor, turn on nitrogen, and react at 25-28℃ for 45 min until the reaction is complete. Filter to remove the liquid. Wash the resin three times with 1 L of DMF solution each time. After washing, add 1 L of 20% Piperidine / DMF solution, react at 25.0-28.0℃ for 30 min, and filter to remove the liquid. Then wash six times with 1 L of DMF solution and dry for later use.

[0045] S13: Prepare 2L of capping solution (DMF:Ac2O:DIEA = 84:10:6) and add it to the reactor. Turn on nitrogen and react at 25~28℃ for 45 min. After the reaction, take a sample and test for ninhydrin. If the result is negative, stop stirring and filter to remove the liquid. Add 1000mL±100mL of DMF to the reactor, stir and wash, then filter to remove the liquid. Repeat the washing operation after coupling twice, for a total of three washings.

[0046] S14: The peptide resin was washed twice with methyl ether, 1 L each time, and the liquid was removed by filtration; washed twice with dichloromethane, 1 L each time, and the liquid was removed by filtration; washed twice with methyl ether, 1 L each time, and the liquid was removed by filtration. The wet product was vacuum dried for more than 12 hours to obtain 940 g of the fully protected peptide resin Ac-Lys(Boc)-Glu(OtBu)-Pro-Pro-Pro-Lys(Boc)-Arg(Pbf)-Pro-Ala-Asp(OtBu)-Linker-AM Resin, namely A6PR2498A.

[0047] S15-1: Deprotection cleavage. Weigh 200g of A6PR2498A and add it to 2L of cleavage solution with a ratio of TFA:Tis:H2O = 95:2.5:2.5. React in a shaker at 30℃ for 2 hours. After filtration, precipitate the solid by sedimentation with 20L of methyl ether. Separate the supernatant by centrifugation, and then wash with methyl ether and centrifuge three times, 5L each time. Dry the solid under vacuum to obtain 89.6g of the fully protected polypeptide Ac-Lys-Glu-Pro-Pro-Pro-Lys-Arg-Pro-Ala-Asp-NH2, i.e., A6PR2498B, with a crude product yield of 92.5%.

[0048] S15-2: Deprotection cleavage. Weigh 200g of A6PR2498A and add it to 2L of cleavage solution. The cleavage solution ratio is TFA:Tis:EDT:PhOH:H2O = 87.5:5:2.5:2.5:2.5. Incubate at 30℃ on a shaker for 2 hours. After filtration, precipitate the solid by sedimentation with 20L of methyl ether. Centrifuge to separate the supernatant, then wash with methyl ether and centrifuge 3 times, 5L each time. Dry the solid under vacuum to obtain 81.6g of the fully protected polypeptide Ac-Lys-Glu-Pro-Pro-Pro-Lys-Arg-Pro-Ala-Asp-NH2, i.e., A6PR2498B, with a crude product yield of 84.3%.

[0049] S15-3: Deprotection cleavage. Weigh 200g of A6PR2498A and add it to 2L of cleavage solution. The cleavage solution ratio is TFA:Tis:H2O:DODT = 95:2.5:2.5 + 8 eq DODT. React in a shaker at 30℃ for 2h. After filtration, the mother liquor is precipitated with 20L of methyl ether to obtain the solid. Centrifuge to separate the supernatant, and then wash with methyl ether and centrifuge 3 times, 5L each time. Dry the solid under vacuum to obtain 74.9g of the fully protected polypeptide Ac-Lys-Glu-Pro-Pro-Pro-Lys-Arg-Pro-Ala-Asp-NH2, i.e., A6PR2498B, with a crude product yield of 77.3%.

[0050] S16: Preferably, we use A6PR2498B obtained under conditions S15-1 for cyclization. Weigh 30g of A6PR2498B and dissolve it in 10L of purified water. Then add 17.9g of DIEA (7eq), and maintain the temperature in a water bath at 30℃, designating this as solution A. Weigh 7.7g of glutaric acid bis(succinimide) ester (1.2eq) and dissolve it in 5L of ACN, designating this as solution B. Slowly add solution B to solution A. The reaction is complete in 10 minutes. TFA is added to the reaction solution to quench the reaction, followed by rotary evaporation to remove all ACN. The concentrated solution is purified by reversed-phase C18 preparative chromatography, and lyophilized to obtain pure product AC-Lys-Glu-Pro-Pro-Pro-Lys-Arg-Pro-Ala-Asp-NH2 (Lys2 & Lys7 by Glutaric Acid), also referred to below as PR2498, with a purity of 98.36%.

[0051] See the high-performance liquid chromatogram and mass spectrum of the peptide compound. Figure 1 and Figure 2 .

[0052] Test Example 1: Cytotoxicity Test of Stapler Peptide Compound HFF-1 cells were seeded into 96-well plates, with 200 μL of culture medium (approximately 10,000 cells / well) added to each well to ensure uniform cell density. The plates were incubated overnight at 37°C with 5% CO2 to allow cell adhesion. After 24 hours, the culture medium was aspirated, and 200 μL of DMEM medium containing different concentrations of the stapling peptide compound PR2498 (0 μM, 1 μM, 5 μM, 10 μM, 50 μM, 100 μM, 500 μM) was added. The 96-well plates were then returned to the incubator for further incubation. After 24 hours, the absorbance was measured at 490 nm using the MTT assay with a microplate reader. A control group (containing only culture medium and no cells) was also included. Cell viability was calculated as (experimental group absorbance - control group absorbance / control group absorbance - control group absorbance) × 100% to determine the cytotoxicity of the stapling peptide compound and its safe concentration range.

[0053] See results Figure 3 ,according to Figure 3 It can be seen that the epithelial cells treated with different concentrations of stapling peptide compound did not show a significant decrease in cell viability. At a concentration of 500 μM, the cell viability remained at 97% relative to the control group, confirming that the stapling peptide compound has extremely low cytotoxicity.

[0054] Test Example 2: Serum Stability Test of Staple Peptide Compounds The lyophilized staple peptide compound PR2498 was directly dissolved in 10% fetal bovine serum to a final concentration of 1 mg / mL. After sterile filtration through a 0.22 μM filter, the solution was placed in a sterile container and incubated at 37°C. Samples were taken at predetermined time points (0 min, 15 min, 30 min, 1 h, 2 h, 4 h, 8 h, 24 h, 48 h). The residue of the staple peptide compound was determined using analytical high-performance liquid chromatography (HPLC), with the content at time 0 as 100%. The decomposition ratio was calculated by comparing the peak areas at different time points. The calculation formula was: Relative content = (Peak area of ​​test sample / Peak area of ​​test sample at time 0) × 100%. A curve of residue percentage versus time was plotted, and the half-life was calculated.

[0055] See results Figure 4 ,according to Figure 4 It can be seen that the staple peptide compound PR2498 is nearly completely degraded in 10% serum after 48 hours, and its half-life is calculated to be about 11.5 hours based on the degradation curve.

[0056] Test Example 3: Regulation of gene expression of various types of collagen by staple peptide compounds HFF-1 fibroblasts in logarithmic growth phase were seeded at 50,000 cells / well into 6-well plates. 2 mL of culture medium was added to each well, and the plates were incubated overnight to allow cells to adhere and grow for 24 hours. Cells were then divided into groups: a 10 μM DMEM medium containing the stapling peptide compound PR2498 was used as a control group; a DMEM blank control group and a 100 ng / mL TGF-β1 positive control group were also established.

[0057] After 24 hours of treatment, the culture medium was removed, and RNA was extracted from cells in each well. The Ct value was detected by qRT-PCR to determine the expression level of type I collagen mRNA in HFF-1 cells at different concentrations. The expression level was calculated using the formula: relative RNA expression level = 2ΔΔC(t).

[0058] Plot the results, and express the results as Mean ± SD. The t-test was used for comparisons between groups. P < 0.05 was considered statistically significant (marked * in the figure), and P < 0.01 was considered highly significant (marked ** in the figure).

[0059] The upregulation rate is calculated using the formula: Upregulation rate = (Sample group - Blank control group) / Blank control group * 100%.

[0060] See results Figures 5-12 ,according to Figures 5-12 It can be seen that, compared with the blank control group, the expression levels of various collagen genes in the positive control group were significantly increased after TGF-β1 treatment, proving that the cell model and qPCR system were working normally.

[0061] Compared with the blank control group, treatment with 10 μM stapling peptide compound PR2498 significantly positively regulated the expression of various collagen genes. Specifically, COLⅠ expression was upregulated by 32%, COLⅢ by 49%, COLⅣ by 142%, COLⅤ by 55%, COLⅥ by 25%, COLⅦ by 46%, COLⅩⅥ by 29%, and COLⅩⅧ by 49%.

[0062] 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. A staple peptide compound, characterized in that, Its structural formula is shown below: 。 2. A method for preparing the staple peptide compound according to claim 1, characterized in that, include: The stapled peptide compound is formed by stapled two side chain residues of a linear polypeptide; wherein the amino acid sequence of the linear polypeptide is as follows: AC-Lys-Glu-Pro-Pro-Pro-Lys-Arg-Pro-Ala-Asp-NH2.

3. The preparation method according to claim 2, characterized in that, The binding process includes reacting the linear polypeptide with activated glutaric acid in a catalyst system.

4. The preparation method according to claim 3, characterized in that, The activation of the glutaric acid can be achieved by using any of the following substances: bis(succinimide), pentafluorophenyl ester, dicyclohexylcarbodiimide carbonate, hydroxybenzotriazole, and azide.

5. The preparation method according to claim 3, characterized in that, The molar ratio of the linear polypeptide to the activated glutaric acid is 1:(1-1.5).

6. The preparation method according to claim 3, characterized in that, The catalyst includes at least one of DIC, HOBt, and DIEA.

7. The preparation method according to claim 3, characterized in that, The molar ratio of the linear polypeptide to the catalyst is 1:(5-10).

8. The use of the staple peptide compound of claim 1 in the preparation of a promoter that promotes collagen expression.

9. The use of the staple peptide compound of claim 1 in the preparation of anti-wrinkle, anti-aging and / or repairing skin care products.

10. A skincare product, characterized in that, It includes the staple peptide compound as described in claim 1.

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