Application of anti-aging repair peptides in cosmetics

Abstract

The present invention discloses the application of anti-aging repair polypeptides in cosmetics, and specifically relates to the technical field of cosmetics. The anti-aging repair polypeptides of the present invention delay and inhibit the synthesis of interleukin-6 through multiple mechanisms of action, stimulate the synthesis of laminin V and collagen IV and VII, promote the synthesis of collagen I, and inhibit the activity of acetylcholine receptors, calcium, sodium, potassium and other ion channels. These effects together make the skin firm and elastic, reduce wrinkles, sagging skin and uneven skin color, and repair the skin and relax muscles to improve and reduce wrinkles.

Description

Technical Field

[0001] The present invention relates to the technical field of cosmetics, and more specifically, to the application of anti-aging and repairing polypeptides in cosmetics. Background Art

[0002] In addition to aging, light exposure is an important factor leading to skin aging and dullness. The resulting skin problems are usually called photoaging. The clinical manifestations of photoaging include dry and rough skin, pigmentation, wrinkle formation, elasticity changes, capillary dilation, purpura and increased incidence of skin tumors.

[0003] With the development of economy, the demand for skin anti-aging and whitening has gradually increased for both men and women. Therefore, more and more scientific research institutions and skin care companies are committed to the research of whitening and anti-aging products to obtain methods or skin care products that can effectively resist skin aging or repair aging skin. At present, the anti-aging ingredients used more in skin care products include: plant extracts, peptides, bosine, vitamin C, vitamin E, fullerene, retinoids, etc. However, any ingredient has certain limitations. In view of the shortcomings of existing ingredients, many researchers have combined multiple ingredients or modified the molecular structure of existing ingredients to obtain more ideal effects.

[0004] Traditional anti-aging ingredients mainly work through anti-oxidation and moisturizing, but the effect is limited. Therefore, the application of anti-aging repair peptides in cosmetics is proposed. Summary of the invention

[0005] In order to overcome the above defects of the prior art, the present invention provides the use of anti-aging repair polypeptides in cosmetics, which delay and inhibit the synthesis of interleukin-6 through multiple mechanisms of action, stimulate the synthesis of laminin V and collagen IV and VII, promote the synthesis of collagen I, and inhibit the activity of acetylcholine receptors, calcium, sodium, potassium and other ion channels. These effects together make the skin firm and elastic, reduce wrinkles, sagging skin and uneven skin color, and repair the skin and relax muscles to improve and reduce wrinkles, so as to solve the problems raised in the above background technology.

[0006] To achieve the above-mentioned object, the present invention provides the following technical scheme: an anti-aging repair polypeptide, whose amino acid sequence is: Palmitoyl-Gly-Gln-Pro-Arg-Gly-Pro-Hyp-Met-Pro-D-Phe-Arg-D-Trp-Phe-Lys-Pro-Val-Arg-Asp-His-Ala-Arg-CONH2.

[0007] A method for synthesizing an anti-aging repair polypeptide, for synthesizing an anti-aging repair polypeptide, comprises the following steps:

[0008] S1: Calculate the weight of raw materials: Calculate the weight of each raw material according to the weight of the target peptide to ensure that the weight of each raw material is calculated accurately;

[0009] S2: Soaking RinkAmideAM resin: Soak the resin in DCM (dichloromethane) for 2 hours to allow the resin to fully swell and prepare for subsequent reactions;

[0010] S3: Washing RinkAmideAM resin: By washing with DMF (N,N-dimethylformamide) and draining, unreacted reagents and impurities can be removed while maintaining the activity of the resin. Repeat this four times and drain the resin;

[0011] S4: Adding the C-terminal amino acid: Fmoc-Arg(pbf)-OH (where "pbf" is a protecting group) was added to the resin and reacted at 30°C for 2 hours to construct the starting part of the polypeptide chain;

[0012] S5: Blocking unreacted groups: Use a mixture of methanol and DIEA (N,N-diisopropylethylamine) to block unreacted amino groups, react for half an hour, then wash with DMF 4 times and drain.

[0013] S6: Removal of Fmoc protecting group: The Fmoc protecting group was removed by using 20% ​​piperidine solution. After removal of protection, it was washed with DMF four times and then dried. A small amount of resin was taken for detection by the ninhydrin method. If the resin had color, it indicated that the deprotection was successful.

[0014] S7: Weigh Fmoc-Ala-OH (the second amino acid at the C-terminus) + HOBT + DIC and add them to the reactor, then place the reactor in a shaker at 30°C for 1 hour;

[0015] S8: Take a small amount of resin for testing using the ninhydrin method. If the resin has color, it means that the condensation is incomplete and the reaction should be continued. If the resin is colorless, it means that the reaction is complete. After the reaction is complete, wash the resin four times with DMF and then drain it;

[0016] S9: Add a certain amount of 20% piperidine to the reactor and shake it on a decolorizing shaker for 20 minutes to remove the Fmoc protecting group on the resin. After deprotection, wash it with DMF four times and then drain it to check whether the protection is removed;

[0017] S10: Take a small amount of resin and test it by ninhydrin method. If the resin has color, it means the deprotection is successful.

[0018] S11: Add the next amino acid: Repeat S7-S10 to add the remaining amino acids in sequence, making sure that the previous amino acid has been correctly connected and protected each time until the last amino acid is synthesized;

[0019] S12: Weigh Palmitoyl+HOBT+DIC and add them into the reactor, then place the reactor in a shaker at 30° C. to react for 1 hour;

[0020] S13: Removal of protecting groups and cleavage of polypeptide: Use a cleavage agent to cleave the polypeptide from the resin and remove all protecting groups to obtain a crude polypeptide;

[0021] S14: Purification of peptides: Purify the peptides by HPLC to ensure that their purity meets the required standards;

[0022] S15: Peptide testing: The molecular weight and purity of the peptide are tested by mass spectrometry and HPLC to ensure the success of the synthesis and the quality of the product.

[0023] Preferably, the mass ratio of the components in S5 is methanol:DIEA:DCM=1:1:2.

[0024] Preferably, the mass percentage of the 20% piperidine and DMF components in S9 is piperidine:DMF=1:4.

[0025] A cosmetic composition containing an anti-aging repair polypeptide, characterized in that it comprises an anti-aging repair polypeptide as claimed in claim 1, and the composition further comprises a cosmetically acceptable carrier and excipients, wherein the content of the anti-aging repair polypeptide in the composition ranges from 10 to 1000 ppm.

[0026] Preferably, the composition can be incorporated into cosmetics such as facial masks, essences, creams, cleansers, toners, gels, lotions, gel lotions, gel essences, and the like.

[0027] Technical effects and advantages of the present invention:

[0028] The present invention delays and inhibits the synthesis of interleukin-6 through multiple mechanisms by synthesizing anti-aging repair polypeptides. This cytokine plays a key role in inflammatory reactions. Therefore, by inhibiting its synthesis, inflammatory reactions and glycosylation damage can be effectively alleviated. Anti-aging repair polypeptides also stimulate the synthesis of laminin V and collagen IV and VII. These proteins are important components of the skin. They make the skin firm and elastic, and reduce wrinkles, sagging skin and uneven skin color. Anti-aging repair polypeptides can also promote the synthesis of collagen I, which is crucial for repairing skin. Collagen is the main structural protein of the skin, which provides support and elasticity for the skin. By increasing the synthesis of collagen, the overall texture and appearance of the skin can be improved. Anti-aging repair polypeptides can also inhibit the activity of acetylcholine receptors, calcium, sodium, potassium and other ion channels. These ion channels play an important role in muscle contraction and relaxation. By inhibiting their activity, muscles can be relaxed and the appearance of wrinkles can be improved and alleviated.

[0029] In summary, anti-aging repair peptides use multiple mechanisms to delay and inhibit the synthesis of interleukin-6, stimulate the synthesis of laminin V and collagen IV and VII, promote the synthesis of collagen I, and inhibit the activity of acetylcholine receptors, calcium, sodium, potassium and other ion channels. These effects work together to make the skin firm and elastic, reduce wrinkles, sagging skin and uneven skin tone, while repairing the skin and relaxing muscles to improve and reduce wrinkles. DETAILED DESCRIPTION

[0030] Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in the field without making any creative work shall fall within the scope of protection of the present invention.

[0031] An anti-aging repair polypeptide, whose amino acid sequence is: Palmitoyl-Gly-Gln-Pro-Arg-Gly-Pro-Hyp-Met-Pro-D-Phe-Arg-D-Trp-Phe-Lys-Pro-Val-Arg-Asp-His-Ala-Arg-CONH2.

[0032] A method for synthesizing an anti-aging repair polypeptide, for synthesizing an anti-aging repair polypeptide, comprises the following steps:

[0033] S1: Calculate the weight of raw materials: Calculate the weight of each raw material according to the weight of the target peptide to ensure that the weight of each raw material is calculated accurately;

[0034] S2: Soaking RinkAmideAM resin: Soak the resin in DCM (dichloromethane) for 2 hours to allow the resin to fully swell and prepare for subsequent reactions;

[0035] S3: Washing RinkAmideAM resin: By washing with DMF (N,N-dimethylformamide) and draining, unreacted reagents and impurities can be removed while maintaining the activity of the resin. Repeat this four times and drain the resin;

[0036] S4: Adding C-terminal amino acid: Fmoc-Arg(pbf)-OH (where "pbf" is a protecting group) was added to the resin and reacted at 30°C for 2 hours to construct the starting part of the polypeptide chain to synthesize Arg(pbf)-Rink Amide AM resin;

[0037] S5: Block unreacted groups: Use a mixture of methanol and DIEA (N,N-diisopropylethylamine) to block unreacted amino groups, methanol: DIEA: DCM = 1:1:2, react for half an hour, then wash with DMF 4 times and drain.

[0038] S6: Removal of Fmoc protecting group: The Fmoc protecting group was removed by using 20% ​​piperidine solution. After removal of protection, it was washed with DMF four times and then dried. A small amount of resin was taken for detection by the ninhydrin method. If the resin had color, it indicated that the deprotection was successful.

[0039] S7: Weigh Fmoc-Ala-OH (the second amino acid at the C-terminus) + HOBT + DIC and add them to the reactor, then place the reactor in a shaker at 30°C for 1 hour;

[0040] S8: Take a small amount of resin for testing using the ninhydrin method. If the resin has color, it means that the condensation is incomplete and the reaction should be continued. If the resin is colorless, it means that the reaction is complete. After the reaction is complete, wash the resin four times with DMF and then drain it;

[0041] S9: Add a certain amount of 20% piperidine to the reactor, piperidine:DMF=1:4, and shake on a decolorizing shaker for 20 minutes to remove the Fmoc protecting group on the resin. After deprotection, wash it with DMF four times, then drain it to check whether the protection is removed;

[0042] S10: Take a small amount of resin and test it by ninhydrin method. If the resin has color, it means the deprotection is successful, and Ala-Arg(pbf)-Rink Amide AM resin is synthesized;

[0043] S11: Add the next amino acid: Repeat S7-S10 to add the remaining amino acids in sequence, making sure that the previous amino acid has been correctly connected and protected each time until the last amino acid is synthesized;

[0044] Specifically, Fmoc-His(Boc)-OH (the third amino acid at the C-terminus) + HOBT + DIC were weighed and added to the reactor, and then the reactor was placed in a shaker at 30°C for reaction for 1 hour, and a small amount of resin was taken for detection by the ninhydrin method. If the resin has color, it means that the condensation is incomplete, and the reaction is continued. If the resin is colorless, it means that the reaction is complete. After the reaction is complete, the resin is washed four times with DMF, and then drained. A certain amount of 20% piperidine was added to the reactor, and piperidine:DMF=1:4. The resin was placed on a decolorization shaker and shaken for 20 minutes to remove the Fmoc protecting group on the resin. After the protection is removed, it was washed four times with DMF, and then drained to detect whether the protection is removed. A small amount of resin was taken for detection by the ninhydrin method. If the resin has color, it means that the deprotection is successful, and His(Boc)-Ala-Arg(Pbf)-Rink Amide AM resin is synthesized;

[0045] Weigh Fmoc-Asp(OtBu)-OH (the fourth amino acid at the C-terminus) + HOBT + DIC and add them to the reactor, then place the reactor in a shaker at 30°C for reaction for 1 hour, take a small amount of resin for detection, and use the ninhydrin method to detect. If the resin has color, it means that the condensation is incomplete, and the reaction continues. If the resin is colorless, it means that the reaction is complete. After the reaction is complete, wash the resin four times with DMF, then drain it, add a certain amount of 20% piperidine to the reactor, piperidine: DMF = 1:4, and shake it on a decolorization shaker for 20 minutes to remove the Fmoc protecting group on the resin. After deprotection, wash it four times with DMF, then drain it to detect whether the protection is removed, take a small amount of resin and use the ninhydrin method to detect. If the resin has color, it means that the deprotection is successful, and Asp(OtBu)-His(Boc)-Ala-Arg(Pbf)-Rink Amide AM resin is synthesized;

[0046] Weigh Fmoc-Arg(Pbf)-OH (the fifth amino acid at the C-terminus) + HOBT + DIC and add them to the reactor, then place the reactor in a shaker at 30°C for reaction for 1 hour, take a small amount of resin for detection, and use the ninhydrin method to detect. If the resin has color, it means that the condensation is incomplete, and the reaction continues. If the resin is colorless, it means that the reaction is complete. After the reaction is complete, wash the resin four times with DMF, then drain it, add a certain amount of 20% piperidine to the reactor, piperidine: DMF = 1:4, and shake it on a decolorization shaker for 20 minutes to remove the Fmoc protecting group on the resin. After deprotection, wash it four times with DMF, then drain it to detect whether the protection is removed, take a small amount of resin and use the ninhydrin method to detect. If the resin has color, it means that the deprotection is successful, and Arg(Pbf)-Asp(OtBu)-His(Boc)-Ala-Arg(Pbf)-Rink Amide AM resin is synthesized;

[0047] Weigh Fmoc-Val-OH (the sixth amino acid at the C-terminus) + HOBT + DIC and add them to the reactor, then place the reactor in a shaker at 30°C for reaction for 1 hour, take a small amount of resin for detection, and use the ninhydrin method to detect. If the resin has color, it means that the condensation is incomplete, and the reaction continues. If the resin is colorless, it means that the reaction is complete. After the reaction is complete, wash the resin four times with DMF, then drain it, add a certain amount of 20% piperidine to the reactor, piperidine: DMF = 1:4, and shake it on a decolorization shaker for 20 minutes to remove the Fmoc protecting group on the resin. After deprotection, wash it four times with DMF, then drain it to detect whether the protection is removed, take a small amount of resin and use the ninhydrin method to detect. If the resin has color, it means that the deprotection is successful, and the Val-Arg(Pbf)-Asp(OtBu)-His(Boc)-Ala-Arg(Pbf)-Rink Amide AM resin is synthesized;

[0048] Weigh Fmoc-Pro-OH (the seventh amino acid at the C-terminus) + HOBT + DIC and add them to the reactor, then place the reactor in a shaker at 30°C for reaction for 1 hour, take a small amount of resin for detection, and use the ninhydrin method to detect. If the resin has color, it means that the condensation is incomplete, and the reaction continues. If the resin is colorless, it means that the reaction is complete. After the reaction is complete, wash the resin four times with DMF, then drain it, add a certain amount of 20% piperidine to the reactor, piperidine: DMF = 1:4, and shake it on a decolorization shaker for 20 minutes to remove the Fmoc protecting group on the resin. After deprotection, wash it four times with DMF, then drain it to detect whether the protection is removed, take a small amount of resin and use the ninhydrin method to detect. If the resin has color, it means that the deprotection is successful, and Pro-Val-Arg(Pbf)-Asp(OtBu)-His(Boc)-Ala-Arg(Pbf)-Rink Amide AM resin is synthesized;

[0049] Weigh Fmoc-Lys(Boc)-OH (the eighth amino acid at the C-terminus) + HOBT + DIC and add them to the reactor, then place the reactor in a shaker at 30°C for reaction for 1 hour, take a small amount of resin for detection, and use the ninhydrin method to detect. If the resin has color, it means that the condensation is incomplete, and the reaction is continued. If the resin is colorless, it means that the reaction is complete. After the reaction is complete, wash the resin four times with DMF, then drain it, add a certain amount of 20% piperidine to the reactor, piperidine: DMF = 1:4, and shake it on a decolorization shaker for 20 minutes to remove the Fmoc protecting group on the resin. After deprotection, wash it four times with DMF, then drain it to detect whether the protection is removed, take a small amount of resin and use the ninhydrin method to detect. If the resin has color, it means that the deprotection is successful, and Lys(Boc)-Pro-Val-Arg(Pbf)-Asp(OtBu)-His(Boc)-Ala-Arg(Pbf)-Rink Amide AM resin is synthesized;

[0050] Weigh Fmoc-Phe-OH (the ninth amino acid at the C-terminus) + HOBT + DIC and add them to the reactor, then place the reactor in a shaker at 30°C for reaction for 1 hour, take a small amount of resin for detection, and use the ninhydrin method to detect. If the resin has color, it means that the condensation is incomplete, and the reaction is continued. If the resin is colorless, it means that the reaction is complete. After the reaction is complete, wash the resin four times with DMF, then drain it, add a certain amount of 20% piperidine to the reactor, piperidine: DMF = 1:4, and shake it on a decolorization shaker for 20 minutes to remove the Fmoc protecting group on the resin. After deprotection, wash it four times with DMF, then drain it to detect whether the protection is removed, take a small amount of resin and use the ninhydrin method to detect. If the resin has color, it means that the deprotection is successful, and Phe-Lys(Boc)-Pro-Val-Arg(Pbf)-Asp(OtBu)-His(Boc)-Ala-Arg(Pbf)-Rink Amide AM resin is synthesized;

[0051] Weigh Fmoc-D-Trp(Boc)-OH (the tenth amino acid at the C-terminus) + HOBT + DIC and add them to the reactor. Then place the reactor in a shaker at 30°C for 1 hour. Take a small amount of resin for testing using the ninhydrin method. If the resin has color, it means that the condensation is incomplete. Continue the reaction. If the resin is colorless, it means that the reaction is complete. After the reaction is complete, wash the resin four times with DMF, then drain it, and add a certain amount of 20% piperidine to the reactor. Piperidine:DMF=1:4 , shake on a decolorizing shaker for 20 minutes to remove the Fmoc protecting group on the resin. After deprotection, wash with DMF four times, then drain to detect whether the protection is removed. Take a small amount of resin and detect it by ninhydrin method. If the resin has color, it means that the deprotection is successful, and D-Trp(Boc)-Phe-Lys(Boc)-Pro-Val-Arg(Pbf)-Asp(OtBu)-His(Boc)-Ala-Arg(Pbf)-Rink Amide AM resin is synthesized;

[0052] Weigh Fmoc-Arg(Pbf)-OH (the eleventh amino acid at the C-terminus) + HOBT + DIC and add them to the reactor, then place the reactor in a shaker at 30°C for 1 hour, take a small amount of resin for detection, and use the ninhydrin method to detect. If the resin has color, it means that the condensation is incomplete, and the reaction is continued. If the resin is colorless, it means that the reaction is complete. After the reaction is complete, wash the resin four times with DMF, then drain it, add a certain amount of 20% piperidine to the reactor, piperidine: DMF = 1:4, and place it on a decolorizing shaker. The resin was shaken for 20 minutes to remove the Fmoc protecting group on the resin. After the protection was removed, it was washed with DMF four times, and then dried to check whether the protection was removed. A small amount of resin was taken for detection by the ninhydrin method. If the resin had color, it indicated that the deprotection was successful, and Arg(Pbf)-D-Trp(Boc)-Phe-Lys(Boc)-Pro-Val-Arg(Pbf)-Asp(OtBu)-His(Boc)-Ala-Arg(Pbf)-RinkAmide AM resin was synthesized.

[0053] Weigh Fmoc-D-Phe-OH (the twelfth amino acid at the C-terminus) + HOBT + DIC and add them to the reactor, then place the reactor in a shaker at 30°C for 1 hour, take a small amount of resin for detection, and use the ninhydrin method to detect. If the resin has color, it means that the condensation is incomplete, and continue the reaction. If the resin is colorless, it means that the reaction is complete. After the reaction is complete, wash the resin four times with DMF, then drain it, add a certain amount of 20% piperidine to the reactor, piperidine: DMF = 1:4, and shake it on a decolorizing shaker for 20 min, in order to remove the Fmoc protecting group on the resin, wash it with DMF four times after deprotection, then drain it to detect whether the protection is removed, take a small amount of resin and detect it by ninhydrin method, the resin has color, indicating that the deprotection is successful, and D-Phe-Arg(Pbf)-D-Trp(Boc)-Phe-Lys(Boc)-Pro-Val-Arg(Pbf)-Asp(OtBu)-His(Boc)-Ala-Arg(Pbf)-RinkAmide AM resin is synthesized;

[0054] Weigh Fmoc-Pro-OH (the thirteenth amino acid at the C-terminus) + HOBT + DIC and add them to the reactor, then place the reactor in a shaker at 30°C for 1 hour, take a small amount of resin for detection, and use the ninhydrin method to detect. If the resin has color, it means that the condensation is incomplete, and continue the reaction. If the resin is colorless, it means that the reaction is complete. After the reaction is complete, wash the resin four times with DMF, then drain it, add a certain amount of 20% piperidine to the reactor, piperidine: DMF = 1:4, and shake it on a decolorizing shaker for 20 min, in order to remove the Fmoc protecting group on the resin, wash it with DMF four times after deprotection, then drain it to check whether the protection is removed, take a small amount of resin and test it by ninhydrin method, the resin has color, indicating that the deprotection is successful, and Pro-D-Phe-Arg(Pbf)-D-Trp(Boc)-Phe-Lys(Boc)-Pro-Val-Arg(Pbf)-Asp(OtBu)-His(Boc)-Ala-Arg(Pbf)-Rink Amide AM resin is synthesized;

[0055] Weigh Fmoc-Met-OH (the fourteenth amino acid at the C-terminus) + HOBT + DIC and add them to the reactor, then place the reactor in a shaker at 30°C for 1 hour, take a small amount of resin for detection, and use the ninhydrin method to detect. If the resin has color, it means that the condensation is incomplete, and continue the reaction. If the resin is colorless, it means that the reaction is complete. After the reaction is complete, wash the resin four times with DMF, then drain it, add a certain amount of 20% piperidine to the reactor, piperidine: DMF = 1:4, and shake it on a decolorizing shaker for 20 minutes. n, in order to remove the Fmoc protecting group on the resin, wash it with DMF four times after deprotection, then drain it to check whether the protection is removed, take a small amount of resin and detect it by ninhydrin method, the resin has color, indicating that the deprotection is successful, and Met-Pro-D-Phe-Arg(Pbf)-D-Trp(Boc)-Phe-Lys(Boc)-Pro-Val-Arg(Pbf)-Asp(OtBu)-His(Boc)-Ala-Arg(Pbf)-Rink Amide AM resin is synthesized;

[0056] Weigh Fmoc-Hyp(tBu)-OH (the fifteenth amino acid at the C-terminus) + HOBT + DIC and add them to the reactor, then place the reactor in a shaker at 30°C for 1 hour, take a small amount of resin for detection, and use the ninhydrin method to detect. If the resin has color, it means that the condensation is incomplete, and continue the reaction. If the resin is colorless, it means that the reaction is complete. After the reaction is complete, wash the resin four times with DMF, then drain it, add a certain amount of 20% piperidine to the reactor, piperidine: DMF = 1:4, and shake it on a decolorizing shaker for 20 minutes. In this way, the Fmoc protecting group on the resin is removed. After the protection is removed, it is washed with DMF four times, then dried to detect whether the protection is removed. A small amount of resin is taken for detection by the ninhydrin method. If the resin has color, it means that the deprotection is successful, and Hyp(tBu)-Met-Pro-D-Phe-Arg(Pbf)-D-Trp(Boc)-Phe-Lys(Boc)-Pro-Val-Arg(Pbf)-Asp(OtBu)-His(Boc)-Ala-Arg(Pbf)-Rink Amide AM resin is synthesized;

[0057] Weigh Fmoc-Pro-OH (the sixteenth amino acid at the C-terminus) + HOBT + DIC and add them to the reactor, then place the reactor in a shaker at 30°C for 1 hour, take a small amount of resin for detection, and use the ninhydrin method to detect. If the resin has color, it means that the condensation is incomplete, and continue the reaction. If the resin is colorless, it means that the reaction is complete. After the reaction is complete, wash the resin four times with DMF, then drain it, add a certain amount of 20% piperidine to the reactor, piperidine: DMF = 1:4, and shake it on a decolorizing shaker for 20 minutes to decolorize. Remove the Fmoc protecting group on the resin, wash it with DMF four times after deprotection, then drain it to check whether the protection is removed, take a small amount of resin and detect it by ninhydrin method, the resin has color, indicating that the deprotection is successful, and synthesize Pro-Hyp(tBu)-Met-Pro-D-Phe-Arg(Pbf)-D-Trp(Boc)-Phe-Lys(Boc)-Pro-Val-Arg(Pbf)-Asp(OtBu)-His(Boc)-Ala-Arg(Pbf)-Rink Amide AM resin;

[0058] Weigh Fmoc-Gly-OH (the seventeenth amino acid at the C-terminus) + HOBT + DIC and add them to the reactor, then place the reactor in a shaker at 30°C for 1 hour, take a small amount of resin for detection, and use the ninhydrin method to detect. If the resin has color, it means that the condensation is incomplete, and continue the reaction. If the resin is colorless, it means that the reaction is complete. After the reaction is complete, wash the resin four times with DMF, then drain it, add a certain amount of 20% piperidine to the reactor, piperidine: DMF = 1:4, and shake it on a decolorizing shaker for 20 minutes to remove the resin. The Fmoc protecting group on the lipid was removed and washed four times with DMF, then dried to detect whether the protection was removed. A small amount of resin was taken and tested by the ninhydrin method. The resin had color, indicating that the deprotection was successful. Gly-Pro-Hyp(tBu)-Met-Pro-D-Phe-Arg(Pbf)-D-Trp(Boc)-Phe-Lys(Boc)-Pro-Val-Arg(Pbf)-Asp(OtBu)-His(Boc)-Ala-Arg(Pbf)-Rink Amide AM resin was synthesized;

[0059] Weigh Fmoc-Arg(Pbf)-OH (the eighteenth amino acid at the C-terminus) + HOBT + DIC and add them to the reactor, then place the reactor in a shaker at 30°C for 1 hour, take a small amount of resin for detection, and use the ninhydrin method to detect. If the resin has color, it means that the condensation is incomplete, and continue the reaction. If the resin is colorless, it means that the reaction is complete. After the reaction is complete, wash the resin four times with DMF, then drain it, add a certain amount of 20% piperidine to the reactor, piperidine: DMF = 1:4, and shake it on a decolorizing shaker for 20 minutes to remove the resin. The Fmoc protecting group was removed and washed with DMF four times. The resin was then drained to detect whether the protection was removed. A small amount of resin was tested by the ninhydrin method. The resin had color, indicating that the deprotection was successful. Arg(Pbf)-Gly-Pro-Hyp(tBu)-Met-Pro-D-Phe-Arg(Pbf)-D-Trp(Boc)-Phe-Lys(Boc)-Pro-Val-Arg(Pbf)-Asp(OtBu)-His(Boc)-Ala-Arg(Pbf)-Rink Amide AM resin was synthesized.

[0060] Weigh Fmoc-Pro-OH (the nineteenth amino acid at the C-terminus) + HOBT + DIC and add them to the reactor, then place the reactor in a shaker at 30°C for 1 hour, take a small amount of resin for detection, and use the ninhydrin method to detect. If the resin has color, it means that the condensation is incomplete, and continue the reaction. If the resin is colorless, it means that the reaction is complete. After the reaction is complete, wash the resin four times with DMF, then drain it, add a certain amount of 20% piperidine to the reactor, piperidine: DMF = 1:4, and shake it on a decolorization shaker for 20 minutes to remove the Fmoc on the resin. After the protection group is removed, it is washed with DMF four times, then dried to check whether the protection is removed. A small amount of resin is taken and tested by the ninhydrin method. The resin has color, indicating that the deprotection is successful. Pro-Arg(Pbf)-Gly-Pro-Hyp(tBu)-Met-Pro-D-Phe-Arg(Pbf)-D-Trp(Boc)-Phe-Lys(Boc)-Pro-Val-Arg(Pbf)-Asp(OtBu)-His(Boc)-Ala-Arg(Pbf)-Rink Amide AM resin is synthesized;

[0061] Weigh Fmoc-Gln(Trt)-OH (the 20th amino acid at the C-terminus) + HOBT + DIC and add them to the reactor, then place the reactor in a shaker at 30°C for 1 hour, take a small amount of resin for detection, and use the ninhydrin method to detect. If the resin has color, it means that the condensation is incomplete, and continue the reaction. If the resin is colorless, it means that the reaction is complete. After the reaction is complete, wash the resin four times with DMF, then drain it, add a certain amount of 20% piperidine to the reactor, piperidine: DMF = 1:4, and shake it on a decolorization shaker for 20 minutes to remove the Fmoc protection on the resin. After deprotection, wash with DMF four times, then drain to check whether the protection is removed. Take a small amount of resin and detect it by ninhydrin method. The resin has color, indicating that the deprotection is successful. Gln(Trt)-Pro-Arg(Pbf)-Gly-Pro-Hyp(tBu)-Met-Pro-D-Phe-Arg(Pbf)-D-Trp(Boc)-Phe-Lys(Boc)-Pro-Val-Arg(Pbf)-Asp(OtBu)-His(Boc)-Ala-Arg(Pbf)-Rink Amide AM resin is synthesized;

[0062] Weigh Fmoc-Gly-OH (the 21st amino acid at the C-terminus) + HOBT + DIC and add them to the reactor, then place the reactor in a shaker at 30°C for 1 hour, take a small amount of resin for detection, and use the ninhydrin method to detect. If the resin has color, it means that the condensation is incomplete, and the reaction is continued. If the resin is colorless, it means that the reaction is complete. After the reaction is complete, wash the resin four times with DMF, then drain it, add a certain amount of 20% piperidine to the reactor, piperidine: DMF = 1:4, and shake it on a decolorization shaker for 20 minutes to remove the Fmoc protecting group on the resin. After the protection is completed, wash it with DMF four times, then drain it to check whether the protection is removed. Take a small amount of resin and detect it by ninhydrin method. If the resin has color, it means that the deprotection is successful, and Gly-Gln(Trt)-Pro-Arg(Pbf)-Gly-Pro-Hyp(tBu)-Met-Pro-D-Phe-Arg(Pbf)-D-Trp(Boc)-Phe-Lys(Boc)-Pro-Val-Arg(Pbf)-Asp(OtBu)-His(Boc)-Ala-Arg(Pbf)-Rink Amide AM resin is synthesized;

[0063] S12: Weigh Palmitoyl+HOBT+DIC and add them to the reactor, then place the reactor in a shaker at 30°C for reaction for 1 hour to synthesize Palmitoyl-Gly-Gln(Trt)-Pro-Arg(Pbf)-Gly-Pro-Hyp(tBu)-Met-Pro-D-Phe-Arg(Pbf)-D-Trp(Boc)-Phe-Lys(Boc)-Pro-Val-Arg(Pbf)-Asp(OtBu)-His(Boc)-Ala-Arg(Pbf)-Rink Amide AM resin;

[0064] S13: Removal of protecting groups and cleavage of polypeptide: The polypeptide is cleaved from the resin using a cleavage reagent, and all protecting groups are removed to obtain a crude polypeptide, namely, Palmitoyl-Gly-Gln-Pro-Arg-Gly-Pro-Hyp-Met-Pro-D-Phe-Arg-D-Trp-Phe-Lys-Pro-Val-Arg-Asp-His-Ala-Arg-CONH2;

[0065] S14: Purification of peptides: Purify the peptides by HPLC to ensure that their purity meets the required standards;

[0066] S15: Peptide testing: The molecular weight and purity of the peptide are tested by mass spectrometry and HPLC to ensure the success of the synthesis and the quality of the product.

[0067] Specifically, the polypeptide sequence Palmitoyl-Gly-Gln-Pro-Arg-Gly-Pro-Hyp-Met-Pro-D-Phe-Arg-D-Trp-Phe-Lys-Pro-Val-Arg-Asp-His-Ala-Arg-CONH2 is represented as follows:

[0068] CONH2 is an amide protecting group, Arg is arginine, Ala is alanine, His is histidine, Asp is aspartic acid, Arg is arginine, Val is valine, Pro is proline, Lys is lysine, Phe is phenylalanine, Phe is phenylalanine, D-Trp is D-tryptophan, Arg is arginine, D-Phe is D-phenylalanine, Pro is proline, Met is methionine, Hyp is hydroxyproline, Pro is proline, Gly is glycine, Arg is arginine, Pro is proline, Gln is glutamine, Gly is glycine, and Palmitoyl is palmitoyl, which is specifically a common fatty acid modification used to connect to the amino group of N-terminal glycine.

[0069] A cosmetic composition containing an anti-aging repair polypeptide comprises the above-mentioned anti-aging repair polypeptide, and the composition further comprises a cosmetically acceptable carrier and auxiliary materials, wherein the content of the anti-aging repair polypeptide in the composition ranges from 10 to 1000 ppm.

[0070] The composition can be incorporated into cosmetics such as facial masks, essences, creams, cleansing milk, toners, gels, lotions, gel lotions, gel essences, and the like.

[0071] Example 1: 10 ppm of anti-aging repair polypeptide is mixed with an appropriate amount of water, glycerin, propylene glycol and other cosmetically acceptable carriers to prepare an essence. After use, the essence can effectively improve the firmness and elasticity of the skin and reduce wrinkles and sagging.

[0072] Example 2: 500 ppm of anti-aging repair polypeptide is mixed with a cream base to prepare an anti-aging cream. After use, the cream can deeply nourish the skin, promote the synthesis of collagen, and make the skin smoother and more delicate.

[0073] Example 3: 1000 ppm of anti-aging repair polypeptide is mixed with a facial cleanser formula to prepare a facial cleanser with anti-aging efficacy.

[0074] The present invention delays and inhibits the synthesis of interleukin-6 through multiple mechanisms by synthesizing anti-aging repair polypeptides. This cytokine plays a key role in inflammatory reactions. Therefore, by inhibiting its synthesis, inflammatory reactions and glycosylation damage can be effectively alleviated. Anti-aging repair polypeptides also stimulate the synthesis of laminin V and collagen IV and VII. These proteins are important components of the skin. They make the skin firm and elastic, and reduce wrinkles, sagging skin and uneven skin color. Anti-aging repair polypeptides can also promote the synthesis of collagen I, which is crucial for repairing skin. Collagen is the main structural protein of the skin, which provides support and elasticity for the skin. By increasing the synthesis of collagen, the overall texture and appearance of the skin can be improved. Anti-aging repair polypeptides can also inhibit the activity of acetylcholine receptors, calcium, sodium, potassium and other ion channels. These ion channels play an important role in muscle contraction and relaxation. By inhibiting their activity, muscles can be relaxed and the appearance of wrinkles can be improved and alleviated.

[0075] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. An anti-aging repair polypeptide, characterized in that: Its amino acid sequence is: Palmitoyl-Gly-Gln-Pro-Arg-Gly-Pro-Hyp-Met-Pro-D-Phe-Arg-D-Trp-Phe-Lys-Pro-Val-Arg-Asp-His-Ala-Arg-CONH2.

2. A method for synthesizing an anti-aging repair polypeptide, for synthesizing the anti-aging repair polypeptide according to claim 1, characterized in that: The following steps are involved: S1: Calculate the weight of raw materials: Calculate the weight of each raw material according to the weight of the target peptide to ensure that the weight of each raw material is calculated accurately; S2: Soaking RinkAmideAM resin: Soak the resin in DCM for 2 hours to allow the resin to fully swell and prepare for subsequent reactions; S3: Washing RinkAmideAM resin: By washing with DMF and draining, unreacted reagents and impurities can be removed while maintaining the activity of the resin. Repeat this four times and drain the resin; S4: Add C-terminal amino acid: Add Fmoc-Arg-OH to the resin and react at 30 °C for 2 hours to construct the starting part of the polypeptide chain; S5: Blocking unreacted groups: Use a mixture of methanol and DIEA to block unreacted amino groups, react for half an hour, then wash with DMF 4 times and drain; S6: Removal of Fmoc protecting group: The Fmoc protecting group was removed by using 20% ​​piperidine solution. After removal of protection, the resin was washed four times with DMF and then dried. A small amount of resin was taken for detection by the ninhydrin method. If the resin had color, it indicated that the deprotection was successful. S7: Weigh Fmoc-Ala-OH+HOBT+DIC and add them into the reactor, then place the reactor in a shaker at 30°C for 1 hour; S8: Take a small amount of resin for testing using the ninhydrin method. If the resin has color, it means that the condensation is incomplete and the reaction should be continued. If the resin is colorless, it means that the reaction is complete. After the reaction is complete, wash the resin four times with DMF and then drain it; S9: Add a certain amount of 20% piperidine to the reactor, shake it on a decolorizing shaker for 20 minutes to remove the Fmoc protecting group on the resin, wash it with DMF four times after deprotection, and then drain it to check whether the protection is removed; S10: Take a small amount of resin and test it by ninhydrin method. If the resin has color, it means the deprotection is successful. S11: Add the next amino acid: Repeat S7-S10 to add the remaining amino acids in sequence, making sure that the previous amino acid has been correctly connected and protected each time until the last amino acid is synthesized; S12: Weigh Palmitoyl+HOBT+DIC and add them into the reactor, then place the reactor in a shaker at 30° C. to react for 1 hour; S13: Removal of protecting groups and cleavage of polypeptide: Use a cleavage reagent to cleave the polypeptide from the resin and remove all protecting groups to obtain a crude polypeptide; S14: Purification of peptides: Purify the peptides by HPLC to ensure that their purity reaches the required standard; S15: Peptide testing: The molecular weight and purity of the peptide are tested by mass spectrometry and HPLC to ensure the success of the synthesis and the quality of the product.

3. The method for synthesizing the anti-aging repair polypeptide according to claim 2, characterized in that: The mass ratio of the components in S5 is methanol:DIEA:DCM=1:1:

2.

4. The method for synthesizing the anti-aging repair polypeptide according to claim 3, characterized in that: The specific mass percentage of the 20% piperidine and DMF components in S9 is piperidine:DMF=1:

4.

5. A cosmetic composition containing an anti-aging repair polypeptide, characterized in that: The composition comprises an anti-aging repair polypeptide as claimed in claim 1, and further comprises a cosmetically acceptable carrier and auxiliary materials, wherein the content of the anti-aging repair polypeptide in the composition ranges from 10 to 1000 ppm.

6. The cosmetic composition containing the anti-aging repair polypeptide according to claim 5, characterized in that: The composition can be incorporated into a facial mask, an essence, a cream, a cleanser, a toner, a gel, an emulsion, a gel emulsion, or a gel essence.