Hyaluronic acid modified heptapeptide as well as preparation method and application thereof

Abstract

The invention discloses hyaluronic acid modified heptapeptide as well as a preparation method and application thereof, and relates to the field of cosmetic peptides. The hyaluronic acid modified cosmetic peptide has a structural formula as shown in a formula (I): M-C (I), wherein M represents sodium hyaluronate, the structure of the sodium hyaluronate is shown in the formula (II), and in the formula (II), y is a natural number larger than or equal to 1; c represents a cosmetic peptide, the cosmetic peptide comprises a polypeptide with cosmetic and / or skin care efficacy or a derivative thereof, and the polypeptide comprises a dipeptide, a tripeptide, a tetrapeptide, a pentapeptide, a hexapeptide, a heptapeptide, an octapeptide or a nonapeptide; wherein M-terminal glucuronic acid is in open-loop linkage with amino in the C structure. The hyaluronic acid modified cosmetic peptide provided by the invention not only can maintain the property of sodium hyaluronate, but also can have the functionality of polypeptide, and the stability of the prepared novel compound is obviously improved, so that the hyaluronic acid modified cosmetic peptide has a wide application prospect in the field of cosmetics / skin care products.

Description

Technical Field

[0001] The present invention belongs to the field of cosmetic peptides, and in particular relates to a hyaluronic acid-modified heptapeptide, a preparation method and uses thereof. Background Art

[0002] Hyaluronic acid is a macromolecular polysaccharide first isolated from bovine vitreous humor by Professors Meyer and Palmer of Ophthalmology at Columbia University in 1934. Its aqueous solution is transparent and glassy, ​​hence its name, hyaluronic acid. Hyaluronic acid is a natural biological substance found widely in tissues such as the vitreous humor, skin, umbilical cord, and synovial fluid. Early hyaluronic acid raw materials were primarily isolated and extracted from rooster combs. Due to cost and raw material limitations, this process was not widely adopted. Currently, hyaluronic acid for cosmetic and pharmaceutical use is primarily produced through microbial fermentation. This process is not restricted by animal raw materials, offers low costs, is easily scalable, and produces high product purity, making it easier to control molecular weight. Research in this field began in China in the 1980s and has achieved remarkable results. Hyaluronic acid produced by fermentation in my country is now internationally advanced in both quality and quantity. Hyaluronic acid is a major component of the extracellular matrix and intercellular matrix, acting as a filler between cells and playing a vital role in the morphology, structure, and function of the skin. Hyaluronic acid is increasingly used in cosmetics due to its moisturizing, repairing, and nourishing properties, good skin compatibility, and safety. Due to its biodegradability, biocompatibility, chemical modification capabilities, and in vivo targeting, hyaluronic acid has attracted considerable attention in the field of protein and polypeptide modification. This invention utilizes hyaluronic acid-modified cosmetic peptides and further investigates their synthesis methods and applications. Summary of the Invention

[0003] The present invention aims to provide a hyaluronic acid-modified cosmetic peptide, a preparation method, and its use. The hyaluronic acid-modified cosmetic peptide can maintain the properties of sodium hyaluronate while containing the functionality of a polypeptide. In addition, the stability of the prepared novel compound is significantly improved, and the peptide has broad application prospects in the field of cosmetics / skin care products.

[0004] The technical solutions adopted by the present invention to achieve the above-mentioned purpose are: A hyaluronic acid-modified beauty peptide, the structural formula of the hyaluronic acid-modified beauty peptide is shown in formula (I): MC (I); Wherein, M represents sodium hyaluronate, and its structure is shown in formula (II): (II), where y is a natural number ≥ 1; C represents a beauty peptide, which includes a polypeptide or a derivative thereof having beauty and / or skin care effects, and includes a dipeptide, tripeptide, tetrapeptide, pentapeptide, hexapeptide, heptapeptide, octapeptide, nonapeptide or other polypeptides; The M-terminal glucuronic acid links the amino group in the C structure. The present invention modifies hyaluronic acid with a cosmetic peptide. Using a substitution reaction, sodium hyaluronate and the peptide are covalently bonded to form a novel chemical compound that retains the properties of sodium hyaluronate while also incorporating the functionality of the peptide. Furthermore, the compound exhibits multiple benefits, with significantly enhanced stability and efficacy compared to the original. For example, the compound exhibits superior moisturizing properties; some compounds exhibit significantly enhanced anti-wrinkle and firming, whitening, antioxidant, and anti-glycation properties, as well as improved soothing properties. The present invention covalently couples hyaluronic acid and a cosmetic peptide to produce a hyaluronic acid-modified cosmetic peptide. This direct chemical modification creates a novel chemical structure, fundamentally endowing the compound with superior properties. The preparation method is simple. The present invention also provides its use in cosmetics and beauty products, significantly enhancing the effectiveness of functional cosmetics, such as hydration, anti-aging and firming, whitening, soothing, antioxidant, and anti-glycation benefits, thereby increasing user satisfaction.

[0005] Preferably, y is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12; more preferably, y=1 or 2 or 3.

[0006] Specifically, the compound represented by formula (I) includes the structure represented by the following formula (III): (III); in, The above n is a natural number; The above R is the remaining part of the cosmetic peptide structure after removing the reactive amino group; The above-mentioned beauty peptides include polypeptides or derivatives thereof having beauty and / or skin care effects, and the above-mentioned polypeptides include dipeptides, tripeptides, tetrapeptides, pentapeptides, hexapeptides, heptapeptides, octapeptides, nonapeptides or other polypeptides.

[0007] Preferably, n=0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24. More preferably, n=0 or 1 or 2 or 3.

[0008] Specifically, the dipeptide or its derivative includes one of dipeptide-2 and carnosine; preferably carnosine.

[0009] Specifically, the tripeptide or its derivative includes one of a snake venom peptide, tripeptide-1, tripeptide-5, tripeptide-8, tripeptide-38 and palmitoyl tripeptide-38; preferably tripeptide-1 or tripeptide-8.

[0010] Specifically, the tetrapeptide or its derivative includes one of tetrapeptide-5, tetrapeptide-7, tetrapeptide-9, tetrapeptide-11, tetrapeptide-30 and tetrapeptide-15, or includes a tetrapeptide having an amino acid sequence of H-Asp-Val-Lys-Tyr-OH; preferably tetrapeptide-7 or tetrapeptide-15.

[0011] Specifically, the pentapeptide or its derivative includes one of pentapeptide-4 and myristyl pentapeptide-4; preferably pentapeptide-4.

[0012] Specifically, the hexapeptide or its derivative includes one of hexapeptide-1, hexapeptide-8, hexapeptide-9, hexapeptide-11 and hexapeptide-38, or includes one of the hexapeptides having an amino acid sequence of H-Arg-Arg-Gln-Met-Glu-Glu-NH2, H-Arg-Arg-Gln-D-Met-Glu-Glu-NH2, H-Trp-Phe-Arg-Leu-Ala-His-NH2 and H-Trp-Phe-Arg-D-Leu-Ala-His-NH2.

[0013] Preferably, the hexapeptide or its derivative includes hexapeptide-1, hexapeptide-8, hexapeptide-9, and hexapeptide-11; more preferably, the amino acid sequence is one of the hexapeptides H-Arg-Arg-Gln-Met-Glu-Glu-NH2, H-Arg-Arg-Gln-D-Met-Glu-Glu-NH2, H-Trp-Phe-Arg-Leu-Ala-His-NH2 and H-Trp-Phe-Arg-D-Leu-Ala-His-NH2.

[0014] Specifically, the heptapeptide or its derivatives include a heptapeptide having an amino acid sequence of H-Glu-Glu-Met-Gln-Arg-Arg-Ala-OH or H-Glu-Glu-Met-Gln-Arg-Arg-Ala-NH2.

[0015] Specifically, the octapeptide or its derivative includes an octapeptide having an amino acid sequence of H-Glu-Glu-Met-Gln-Arg-Arg-Ala-Asp-NH2.

[0016] Specifically, the nonapeptide or its derivative includes nonapeptide-1.

[0017] Specifically, the compound shown in (I) above includes one or more of H-Glu-Glu-Met-Gln-Arg-Arg-NH-hyaluronic acid (sodium), hyaluronic acid (sodium)-NH-Phe-Val-Ala-Pro-Phe-Pro-OH, H-His-D-Phe-Arg-NH-hyaluronic acid (sodium), H-His-Ala-Leu-Arg-Phe-Trp-NH-hyaluronic acid (sodium), H-Trp-Phe-Arg-D-Leu-Ala-His-NH-hyaluronic acid (sodium), H-Trp-Phe-Arg-Leu-Ala-His-NH-hyaluronic acid (sodium), H-Arg-Arg-Gln-D-Met-Glu-Glu-NH-hyaluronic acid (sodium) or H-Arg-Arg-Gln-Met-Glu-Glu-NH-hyaluronic acid (sodium).

[0018] Furthermore, the compound shown in (I) above may also include one or more of hyaluronic acid (sodium)-HN-Gly-Gln-Pro-Arg-OH, hyaluronic acid (sodium)-HN-Lys-Thr-Thr-Lys-Ser-OH, H-Lys-Leu-Ala-Lys-Lys-NH-hyaluronic acid (sodium), H-Gly-Pro-Gln-Gly-Pro-Gln-NH-hyaluronic acid (sodium), H-Met-Pro-{D-Phe}-Arg-{D-Trp}-Phe-Lys-Pro-Val-NH-hyaluronic acid (sodium), H-Tyr-Pro-Phe-Phe-NH-hyaluronic acid (sodium), hyaluronic acid (sodium)-NH-Glu-Glu-Met-Gln-Arg-Arg-Ala-OH or hyaluronic acid (sodium)-NH-Asp-Val-Lys-Tyr-OH.

[0019] It should be noted that the hyaluronic acid-modified cosmetic peptides described above may represent a structure in which there is only one reactive site for bonding between hyaluronic acid (sodium) and the cosmetic peptide, or may represent a structure in which there are multiple reactive sites. It should be further noted that the bonding portion comprises a reactive amino group in the cosmetic polypeptide structure and an reactive hydroxyl site in the terminal ring of the hyaluronic acid (sodium) structure that is capable of further polymerization, which reacts.

[0020] More preferably, the compound represented by the above formula (I) includes one or more of the following: A2; A4; A6; B2; B4; B6; D2; D4; D6; E2; E4; E6; F2; F4; F6; G2; G4; G6. The present invention also discloses a method for preparing a hyaluronic acid-modified beauty peptide, comprising: carrying out a condensation reaction between sodium hyaluronate and the beauty peptide to prepare the hyaluronic acid-modified beauty peptide.

[0021] Furthermore, the synthesis route of the above-mentioned hyaluronic acid-modified beauty peptide is as follows: ; Where n is a natural number; Y is Na or H; Peptide=R; R is the remaining part of the cosmetic peptide structure after removing the reactive amino group; X includes H, sodium or potassium; The above-mentioned beauty peptides include polypeptides or derivatives thereof having beauty and / or skin care effects, and the polypeptides include dipeptides, tripeptides, tetrapeptides, pentapeptides, hexapeptides, heptapeptides, octapeptides, nonapeptides or other polypeptides.

[0022] It should be noted that the final form of the hyaluronic acid-modified cosmetic peptide provided by the present invention can be a hyaluronic acid-modified cosmetic peptide, or it can be a hydrochloride, sulfate, phosphate, acetate, sodium salt, potassium salt, trifluoroacetate, maleate or fumarate, etc.

[0023] The preparation method of the hyaluronic acid-modified beauty peptide comprises: using sodium hyaluronate and the beauty peptide to undergo a ring-opening reaction to prepare the hyaluronic acid-modified beauty peptide.

[0024] Specifically, the preparation method of the above-mentioned hyaluronic acid-modified cosmetic peptide comprises the following steps: Add the beauty peptide to the organic solvent, stir to dissolve, then add TFA, stir for 0.5-1h, add DIEA, and then weigh sodium hyaluronate and add; stir and react overnight at 40-50°C, take samples and detect LC-MS, if the reaction is basically complete, add acetic acid, control the temperature in a water bath at 30-45°C, carry out rearrangement reaction for 2-4h, take samples and detect LC-MS, if the reaction is basically complete; then perform reverse chromatography purification to obtain the hyaluronic acid-modified beauty peptide.

[0025] Specifically, the organic solvent is selected from single solvents or mixed solvents such as DMF, N,N-diethylacetamide, DMSO, acetonitrile, nitrogen methyl pyrrolidone (NMP), methanol, ethanol, acetone, THF, dichloromethane, ethyl acetate, dioxane and water; preferably DMSO.

[0026] Specifically, the mass volume ratio of the above-mentioned cosmetic peptide to DMSO is 1g:8-25mL; preferably 1g:10mL.

[0027] Specifically, the mass volume ratio of the above-mentioned beauty peptide to TFA is 1g:0-1.2mL.

[0028] Specifically, the mass ratio of the above-mentioned beauty peptide to DIEA is 1:0.5-5.5; preferably 1:0.8-2.5.

[0029] Specifically, the molar ratio of the above-mentioned cosmetic peptide to hyaluronic acid is 1:2-8; preferably 1:3-5; more preferably 1:4.

[0030] Specifically, the mass volume ratio of the above-mentioned beauty peptide to acetic acid is 1g:4-10mL; preferably 1g:5-9mL; more preferably 1g:8mL.

[0031] Another object of the present invention is to disclose the use of the above-mentioned hyaluronic acid-modified beauty peptide in the preparation of cosmetics and / or skin care products.

[0032] The present invention further discloses the use of the hyaluronic acid-modified beauty peptide in enhancing the moisturizing performance of cosmetics and / or skin care products.

[0033] The present invention also discloses the use of the hyaluronic acid-modified beauty peptide in enhancing the soothing performance of cosmetics and / or skin care products.

[0034] The present invention also discloses the use of the hyaluronic acid-modified beauty peptide in enhancing the anti-wrinkle and firming properties of cosmetics and / or skin care products.

[0035] The present invention also discloses the use of the hyaluronic acid-modified beauty peptide in enhancing the whitening performance of cosmetics and / or skin care products.

[0036] The present invention also discloses the use of the hyaluronic acid-modified beauty peptide in enhancing the anti-glycation performance of cosmetics and / or skin care products.

[0037] The present invention also discloses the use of the hyaluronic acid-modified beauty peptide in enhancing the antioxidant properties of cosmetics and / or skin care products.

[0038] A cosmetic comprising the above-mentioned hyaluronic acid-modified beauty peptide.

[0039] A skin care product comprising the above-mentioned hyaluronic acid-modified beauty peptide.

[0040] The beneficial effects of the present invention include: The present invention uses a beauty peptide to modify hyaluronic acid, and forms a new compound through the Maillard reaction. The compound can maintain the properties of sodium hyaluronate and contain the functionality of the peptide; and its stability and efficacy are significantly enhanced compared to the original efficacy. For example, the compound has more excellent moisturizing properties; the anti-wrinkle and firming properties, whitening properties, antioxidant capacity and anti-glycation properties of some compounds are significantly improved, and the soothing ability is also effectively improved. The hyaluronic acid-modified beauty peptide provided by the present invention is directly modified by chemical modification to form a new chemical structure, which can fundamentally give the compound more excellent properties, and the preparation method is simple. At the same time, the present invention also provides its application in cosmetics and beauty products, which significantly enhances the use effect of functional cosmetics, such as hydration, anti-aging and firming, whitening, soothing, antioxidant and anti-glycation effects, thereby improving user satisfaction.

[0041] Therefore, the present invention provides a hyaluronic acid-modified beauty peptide, a preparation method, and its use. The hyaluronic acid-modified beauty peptide can maintain the properties of sodium hyaluronate while containing the functionality of a polypeptide. In addition, the stability of the prepared novel compound is significantly improved, and it has broad application prospects in the field of cosmetics / skin care products. BRIEF DESCRIPTION OF THE DRAWINGS

[0042] Figure 1 This is the mass spectrum test result of H-Gly-Gln-Pro-Arg-OH prepared in Example 1; Figure 2 This is the liquid chromatography test result of H-Gly-Gln-Pro-Arg-OH prepared in Example 1; Figure 3 This is the mass spectrometry test result of the hyaluronic acid-modified cosmetic peptide prepared in Example 1; Figure 4 This is the liquid chromatography test result of the hyaluronic acid-modified cosmetic peptide prepared in Example 1; Figure 5This is the mass spectrum test result of H-Lys-Thr-Thr-Lys-Ser-OH prepared in Example 2; Figure 6 This is the liquid chromatography test result of H-Lys-Thr-Thr-Lys-Ser-OH prepared in Example 2; Figure 7 This is the liquid chromatography test result of the hyaluronic acid-modified cosmetic peptide prepared in Example 2; Figure 8 This is the mass spectrum test result of the sample at 5.801 min in the liquid chromatography of the hyaluronic acid-modified cosmetic peptide prepared in Example 2; Figure 9 This is the mass spectrum test result of the sample at 6.102 min in the liquid chromatography of the hyaluronic acid-modified cosmetic peptide prepared in Example 2; Figure 10 This is the mass spectrum test result of the sample at 6.558 min in the liquid chromatography of the hyaluronic acid-modified cosmetic peptide prepared in Example 2; Figure 11 This is the mass spectrum test result of H-Lys-Leu-Ala-Lys-Lys-NH2 prepared in Example 3; Figure 12 LC-MS test results of H-Lys-Leu-Ala-Lys-Lys-NH2 prepared in Example 3; Figure 13 This is the mass spectrometry test result of the hyaluronic acid-modified cosmetic peptide prepared in Example 3; Figure 14 This is the liquid chromatography test result of the hyaluronic acid-modified cosmetic peptide prepared in Example 3; Figure 15 This is the mass spectrum test result of H-Glu-Glu-Met-Gln-Arg-Arg-NH2 prepared in Example 4; Figure 16 LC-MS test results of H-Glu-Glu-Met-Gln-Arg-Arg-NH2 prepared in Example 4; Figure 17 This is the mass spectrum test result of the hyaluronic acid-modified cosmetic peptide D2 prepared in Example 4; Figure 18 This is the liquid chromatography test result of the hyaluronic acid-modified cosmetic peptide D2 prepared in Example 4; Figure 19 This is the mass spectrum test result of the hyaluronic acid-modified cosmetic peptide D4 prepared in Example 4; Figure 20 This is the liquid chromatography test result of the hyaluronic acid-modified cosmetic peptide D4 prepared in Example 4; Figure 21 This is the mass spectrum test result of the hyaluronic acid-modified cosmetic peptide D6 prepared in Example 4; Figure 22 This is the liquid chromatography test result of the hyaluronic acid-modified beauty peptide D6 prepared in Example 4; Figure 23 This is the mass spectrometry test result of the hyaluronic acid-modified cosmetic peptide prepared in Example 5; Figure 24 This is the liquid chromatography test result of the hyaluronic acid-modified cosmetic peptide prepared in Example 5; Figure 25 This is the mass spectrum test result of H-Gly-Pro-Gln-Gly-Pro-Gln-NH2 prepared in Example 6; Figure 26 This is the liquid chromatography test result of H-Gly-Pro-Gln-Gly-Pro-Gln-NH2 prepared in Example 6; Figure 27 This is the mass spectrometry test result of the hyaluronic acid-modified cosmetic peptide prepared in Example 6; Figure 28 This is the liquid chromatography test result of the hyaluronic acid-modified cosmetic peptide prepared in Example 6; Figure 29 This is the mass spectrum test result of H-Met-Pro-{D-Phe}-Arg-{D-Trp}-Phe-Lys-Pro-Val-NH2 prepared in Example 7; Figure 30 This is the liquid chromatography test result of H-Met-Pro-{D-Phe}-Arg-{D-Trp}-Phe-Lys-Pro-Val-NH2 prepared in Example 7; Figure 31 This is the mass spectrometry test result of the hyaluronic acid-modified cosmetic peptide prepared in Example 7; Figure 32 This is the liquid chromatography test result of the hyaluronic acid-modified cosmetic peptide prepared in Example 7; Figure 33 This is the mass spectrum test result of H-Phe-Val-Ala-Pro-Phe-Pro-OH prepared in Example 8; Figure 34 This is the LC-MS test result of H-Phe-Val-Ala-Pro-Phe-Pro-OH prepared in Example 8; Figure 35 This is the mass spectrum test result of the hyaluronic acid-modified cosmetic peptide H2 prepared in Example 8; Figure 36This is the liquid chromatography test result of the hyaluronic acid-modified cosmetic peptide H2 prepared in Example 8; Figure 37 This is the mass spectrum test result of the hyaluronic acid-modified cosmetic peptide H4 prepared in Example 8; Figure 38 This is the liquid chromatography test result of the hyaluronic acid-modified cosmetic peptide H4 prepared in Example 8; Figure 39 This is the mass spectrum test result of the hyaluronic acid-modified cosmetic peptide H6 prepared in Example 8; Figure 40 This is the liquid chromatography test result of the hyaluronic acid-modified cosmetic peptide H6 prepared in Example 8; Figure 41 This is the mass spectrum test result of H-His-D-Phe-Arg-NH2 prepared in Example 9; Figure 42 LC-MS test results of H-His-D-Phe-Arg-NH2 prepared in Example 9; Figure 43 This is the mass spectrum test result of the hyaluronic acid-modified cosmetic peptide L2 prepared in Example 9; Figure 44 This is the liquid chromatography test result of the hyaluronic acid-modified cosmetic peptide L2 prepared in Example 9; Figure 45 This is the mass spectrum test result of the hyaluronic acid-modified cosmetic peptide L4 prepared in Example 9; Figure 46 This is the liquid chromatography test result of the hyaluronic acid-modified cosmetic peptide L4 prepared in Example 9; Figure 47 This is the mass spectrum test result of the hyaluronic acid-modified cosmetic peptide L6 prepared in Example 9; Figure 48 This is the liquid chromatography test result of the hyaluronic acid-modified cosmetic peptide L6 prepared in Example 9; Figure 49 This is the mass spectrum test result of H-Tyr-Pro-Phe-Phe-NH2 prepared in Example 10; Figure 50 This is the liquid chromatography test result of H-Tyr-Pro-Phe-Phe-NH2 prepared in Example 10; Figure 51 This is the mass spectrum test result of the hyaluronic acid-modified cosmetic peptide prepared in Example 10; Figure 52 This is the liquid chromatography test result of the hyaluronic acid-modified cosmetic peptide prepared in Example 10; Figure 53The mass spectrometry test results of some products of the hyaluronic acid-modified cosmetic peptide prepared in Example 11 (reaction products of sodium hyaluronate and cosmetic peptide with y=1, y=2, and y=3); Figure 54 HPLC test results of some products (reaction products of sodium hyaluronate and the cosmetic peptide with y=1, y=2, and y=3) of the hyaluronic acid-modified cosmetic peptide prepared in Example 11; Figure 55 The mass spectrometry test results of some products of the hyaluronic acid-modified cosmetic peptide prepared in Example 11 (reaction products of sodium hyaluronate with y=2 and y=3 and the cosmetic peptide); Figure 56 The liquid chromatography test results of some products of the hyaluronic acid-modified cosmetic peptide prepared in Example 11 (reaction products of sodium hyaluronate with y=2 and y=3 and the cosmetic peptide); Figure 57 This is the mass spectrum test result of H-Glu-Glu-Met-Gln-Arg-Arg-Ala-OH prepared in Example 12; Figure 58 This is the liquid chromatography test result of H-Glu-Glu-Met-Gln-Arg-Arg-Ala-OH prepared in Example 12; Figure 59 This is the liquid chromatography test result of the hyaluronic acid-modified cosmetic peptide prepared in Example 12; Figure 60 This is the mass spectrum test result of the sample at 11.577 min in the liquid chromatogram of the hyaluronic acid-modified cosmetic peptide prepared in Example 12; Figure 61 This is the mass spectrum test result of the sample at 12.223 min in the liquid chromatogram of the hyaluronic acid-modified cosmetic peptide prepared in Example 12; Figure 62 This is the mass spectrum test result of H-Asp-Val-Lys-Tyr-OH prepared in Example 13; Figure 63 This is the liquid chromatography test result of H-Asp-Val-Lys-Tyr-OH prepared in Example 13; Figure 64 This is the liquid chromatography test result of the hyaluronic acid-modified cosmetic peptide prepared in Example 13; Figure 65 This is the mass spectrum test result of the sample at 8.226 min in the liquid chromatogram of the hyaluronic acid-modified cosmetic peptide prepared in Example 13; Figure 66This is the mass spectrum test result of the sample at 12.711 min in the liquid chromatogram of the hyaluronic acid-modified cosmetic peptide prepared in Example 13; Figure 67 This is the mass spectrum test result of H-His-Ala-Leu-Arg-Phe-Trp-NH2 prepared in Example 14; Figure 68 This is the liquid chromatography test result of H-His-Ala-Leu-Arg-Phe-Trp-NH2 prepared in Example 14; Figure 69 This is the mass spectrum test result of the hyaluronic acid-modified cosmetic peptide A2 prepared in Example 14; Figure 70 This is the liquid chromatography test result of the hyaluronic acid-modified cosmetic peptide A2 prepared in Example 14; Figure 71 This is the mass spectrum test result of the hyaluronic acid-modified cosmetic peptide A4 prepared in Example 14; Figure 72 This is the liquid chromatography test result of the hyaluronic acid-modified cosmetic peptide A4 prepared in Example 14; Figure 73-1 Partial H NMR spectrum test results of the hyaluronic acid-modified cosmetic peptide A4 prepared in Example 14; Figure 73-2 Partial H NMR spectrum test results of the hyaluronic acid-modified cosmetic peptide A4 prepared in Example 14; Figure 73-3 Partial H NMR spectrum test results of the hyaluronic acid-modified cosmetic peptide A4 prepared in Example 14; Figure 73-4 Partial H NMR spectrum test results of the hyaluronic acid-modified cosmetic peptide A4 prepared in Example 14; Figure 73-5 Partial H NMR spectrum test results of the hyaluronic acid-modified cosmetic peptide A4 prepared in Example 14; Figure 74-1 Partial NMR carbon spectrum test results of the hyaluronic acid-modified cosmetic peptide A4 prepared in Example 14; Figure 74-2 Partial NMR carbon spectrum test results of the hyaluronic acid-modified cosmetic peptide A4 prepared in Example 14; Figure 74-3 Partial NMR carbon spectrum test results of the hyaluronic acid-modified cosmetic peptide A4 prepared in Example 14; Figure 74-4 Partial NMR carbon spectrum test results of the hyaluronic acid-modified cosmetic peptide A4 prepared in Example 14; Figure 74-5Partial NMR carbon spectrum test results of the hyaluronic acid-modified cosmetic peptide A4 prepared in Example 14; Figure 75 This is the two-dimensional cosy map test result of the hyaluronic acid-modified cosmetic peptide A4 prepared in Example 14; Figure 76 This is the mass spectrum test result of the hyaluronic acid-modified cosmetic peptide A6 prepared in Example 14; Figure 77 This is the liquid chromatography test result of the hyaluronic acid-modified cosmetic peptide A6 prepared in Example 14; Figure 78 This is the mass spectrum test result of H-Trp-Phe-Arg-D-Leu-Ala-His-NH2 prepared in Example 27; Figure 79 This is the liquid chromatography test result of H-Trp-Phe-Arg-D-Leu-Ala-His-NH2 prepared in Example 27; Figure 80 This is the liquid chromatography test result of the hyaluronic acid-modified cosmetic peptide E2 prepared in Example 27; Figure 81 This is the mass spectrum test result of the hyaluronic acid-modified cosmetic peptide E2 prepared in Example 27; Figure 82 This is the mass spectrum test result of the hyaluronic acid-modified cosmetic peptide E4 prepared in Example 27; Figure 83 This is the liquid chromatography test result of the hyaluronic acid-modified cosmetic peptide E4 prepared in Example 27; Figure 84 This is the mass spectrum test result of the hyaluronic acid-modified cosmetic peptide E6 prepared in Example 27; Figure 85 This is the liquid chromatography test result of the hyaluronic acid-modified cosmetic peptide E6 prepared in Example 27; Figure 86 This is the mass spectrum test result of H-Trp-Phe-Arg-Leu-Ala-His-NH2 prepared in Example 28; Figure 87 This is the liquid chromatography test result of H-Trp-Phe-Arg-Leu-Ala-His-NH2 prepared in Example 28; Figure 88 This is the mass spectrum test result of the hyaluronic acid-modified cosmetic peptide B2 prepared in Example 28; Figure 89 This is the liquid chromatography test result of the hyaluronic acid-modified cosmetic peptide B2 prepared in Example 28; Figure 90This is the liquid chromatography test result of the hyaluronic acid-modified cosmetic peptide B4 prepared in Example 28; Figure 91 This is the liquid chromatography test result of the hyaluronic acid-modified cosmetic peptide B4 prepared in Example 28; Figure 92 This is the mass spectrum test result of the hyaluronic acid-modified cosmetic peptide B6 prepared in Example 28; Figure 93 This is the liquid chromatography test result of the hyaluronic acid-modified cosmetic peptide B6 prepared in Example 28; Figure 94 This is the mass spectrum test result of H-Arg-Arg-Gln-D-Met-Glu-Glu-NH2 prepared in Example 29; Figure 95 This is the liquid chromatography test result of H-Arg-Arg-Gln-D-Met-Glu-Glu-NH2 prepared in Example 29; Figure 96 This is the liquid chromatography test result of the hyaluronic acid-modified cosmetic peptide G2 prepared in Example 29; Figure 97 This is the liquid chromatography test result of the hyaluronic acid-modified cosmetic peptide G2 prepared in Example 29; Figure 98 This is the liquid chromatography test result of the hyaluronic acid-modified cosmetic peptide G4 prepared in Example 29; Figure 99 This is the mass spectrum test result of the hyaluronic acid-modified cosmetic peptide G4 prepared in Example 29; Figure 100 This is the mass spectrum test result of the hyaluronic acid-modified cosmetic peptide G6 prepared in Example 29; Figure 101 This is the liquid chromatography test result of the hyaluronic acid-modified cosmetic peptide G6 prepared in Example 29; Figure 102 This is the mass spectrum test result of H-Arg-Arg-Gln-Met-Glu-Glu-NH2 prepared in Example 30; Figure 103 This is the liquid chromatography test result of H-Arg-Arg-Gln-Met-Glu-Glu-NH2 prepared in Example 30; Figure 104 This is the liquid chromatography test result of the hyaluronic acid-modified cosmetic peptide F2 prepared in Example 30; Figure 105 This is the liquid chromatography test result of the hyaluronic acid-modified cosmetic peptide F2 prepared in Example 30; Figure 106This is the liquid chromatography test result of the hyaluronic acid-modified cosmetic peptide F4 prepared in Example 30; Figure 107 This is the mass spectrum test result of the hyaluronic acid-modified cosmetic peptide F4 prepared in Example 30; Figure 108 This is the mass spectrum test result of the hyaluronic acid-modified cosmetic peptide F6 prepared in Example 30; Figure 109 This is the liquid chromatography test result of the hyaluronic acid-modified cosmetic peptide F6 prepared in Example 30. DETAILED DESCRIPTION

[0043] In order to make the purpose, technical solutions and advantages of the present invention more clear, the technical solutions of the present invention are further described in detail below in conjunction with specific embodiments: It should be noted that the hyaluronic acid used in Examples 1 to 30 of the present invention is hydrolyzed sodium hyaluronate, purchased from Shandong Baolijia Biotechnology Co., Ltd., which is a mixture. The main components include disaccharide sodium hyaluronate, tetrasaccharide sodium hyaluronate, and hexasaccharide sodium hyaluronate, with a mass ratio of 1:5:3.

[0044] Example 1: A method for synthesizing H-Gly-Gln-Pro-Arg-OH, comprising: 3.13 mmol Wang resin was placed in a 125 mL solid-phase synthesis reactor, and 7.5 mmol amino acid Fmoc-Arg(Pbf)-OH was added. 15 mL of dichloromethane (DCM) was added, followed by pyridine (2.01 mL) and DBU (1.78 mL). The reaction was carried out at 25°C for 3 h, filtered, washed with DMF solution three times, 15 mL each time, and 15 mL of blocking solution (the blocking solution contained Ac2O, DMF, and DIEA, and the mass ratio of Ac2O:DMF:DIEA was 10:84:6) was added and reacted for 15 min. The resin was filtered and washed with dichloromethane twice, 15 mL each time, methanol twice, and DMF twice, 15 mL each time. 15 mL of 20% Pip / DMF (v / v) solution was added and stirred for 30 min. The resin was filtered to remove the deprotection solution, and then washed with DMF six times, 15 mL each time, and dried for use.

[0045] Take 5mmol Fmoc-Pro-OH.H2O and 5mmol HOBt in a 50mL beaker, cool to 5℃, add 5mL DMF and 0.8mL DIC and let it react for 15min. Then add the solution in the 50mL beaker to the above 125mL solid-phase synthesis reactor and stir to react for 1.5h. The reaction is completed. Wash the resin with DMF solution 3 times, 15mL each time. After washing, proceed to the next step of reaction. Add 15mL of 20% Pip / DMF (v / v) solution, stir and react for 30min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 15mL each time, and drain and set aside.

[0046] Take 7.5mmol Fmoc-Gln(Trt)-OH and 7.5mmol HOBt in a 50mL beaker, cool to 5℃, add 5mL DMF and 1.2mL DIC and let it react for 15min. Then add the solution in the 50mL beaker to the above 125mL solid phase synthesis reactor and stir to react for 1.5h. The reaction is completed. Wash the resin with DMF solution 3 times, 15mL each time. After washing, proceed to the next step of reaction. Add 15mL of 20% Pip / DMF (v / v) solution, stir and react for 30min, filter to remove the deprotection solution, and then wash 6 times with 15mL of DMF solution and dry it for use.

[0047] 7.5 mmol Fmoc-Gly-OH and 7.5 mmol HOBt were placed in a 50 mL beaker and cooled to 5°C. 5 mL DMF and 1.2 mL DIC were added and the mixture was allowed to react for 15 min. The solution in the 50 mL beaker was then added to the 125 mL solid-phase synthesis reactor and stirred for 1.5 h. The reaction was completed. The resin was washed with DMF solution three times, 15 mL each time. After washing, the resin was subjected to the next step. 15 mL of 20% Pip / DMF (v / v) solution was added and stirred for 30 min. The deprotection solution was removed by filtration and then washed with DMF solution six times, 15 mL each time, methanol twice, DCM twice, and methanol twice. The resin was dried under vacuum to obtain the peptide resin H-AA1-AA2-AA3-AA4 -Wang-resin, where AA1 is Gly; AA2 is Gln(Trt); AA3 is Pro; and AA4 is Arg(Pbf). The peptide resin was cleaved with TFA / Tis / H2O (volume ratio of TFA, Tis and H2O was 90:5:5) in an amount of 30 mL for 2.5 h. The cleavage solution was added to 300 mL of ether (5°C) solution to precipitate a white solid. After centrifugation, H-Gly-Gln-Pro-Arg-OH was obtained. Its mass spectrum ( Figure 1 ) and high performance liquid chromatography ( Figure 2)like Figure 1-2 shown.

[0048] The preparation method of hyaluronic acid-modified cosmetic peptide comprises the following steps: Weigh 0.46g H-Gly-Gln-Pro-Arg-OH.2TFA, add 5mL DMSO and stir to dissolve, add 0.785g DIEA, and the tetrapeptide precipitates. Then weigh 2.1g hyaluronic acid, control the temperature at 45℃ in a water bath, stir for 15min, and the reaction solution becomes clear. Continue to control the temperature at 45℃ and react overnight. Take a sample and test LC-MS. If the reaction is basically complete, add 4mL acetic acid, control the temperature at 35℃ in a water bath, and carry out the rearrangement reaction for 3h. Take a sample and test LC-MS. If the reaction is basically complete, then perform reverse chromatography purification. Purification conditions: Dissolution: dilute 0.5 g of crude product with 100 mL of H2O; Packing: 21.2*250mm, 10-120, C18; Flow rate: 10mL / min; Wavelength: 220nm; Mobile phase: A: 1% HAc; B: ACN; Balance: A:B=100:0, balance 10 min, flow rate: 10 mL / min; Sample loading: flow rate: 10 mL / min; Elution: 0-10% B 60 min; Column cleaning: 80% ACN cleaning to baseline equilibrium; Qualified products were collected to obtain hyaluronic acid modified beauty peptides, and their mass spectrometry and HPLC characterization results were as follows: Figure 3 and Figure 4 shown.

[0049] Example 2: A method for synthesizing H-Lys-Thr-Thr-Lys-Ser-OH, comprising: 8.75 mmol of CTC resin was placed in a 250 mL solid-phase synthesis reactor, and 17.5 mmol of amino acid Fmoc-Ser(tBu)-OH was added. 75 mL of dichloromethane was added, and the reaction was continued at 25°C for 3 h after adding 7.0 mL of DIEA. 10 mL of methanol was added and the reaction was continued for 5 min. The resin was filtered and washed twice with dichloromethane, 75 mL each time; twice with methanol, 75 mL each time; and twice with DMF, 75 mL each time. 75 mL of 20% Pip / DMF (v / v) solution was added and the reaction was stirred for 30 min. The resin was filtered to remove the deprotection solution, and then washed 6 times with DMF solution, 75 mL each time, and dried for use.

[0050] Take 21mmol Fmoc-Lys(Boc)-OH and 21mmol HOBt in a 100mL beaker, cool to 5℃, add 50mL of DMF solution and 3.2mL of DIC, and let it react for 15min. Then add the solution in the 100mL beaker to the above 250mL solid-phase synthesis reactor and stir to react for 1.5h. The reaction is complete. Wash the resin with DMF solution 3 times, 75mL each time. After washing, proceed to the next step of reaction. Add 75mL of 20% Pip / DMF (v / v) solution, stir and react for 30min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 65mL each time, and drain for use.

[0051] Take 21mmol Fmoc-Thr(tBu)-OH and 21mmol HOBt in a 100mL beaker, cool to 5℃, add 50mL of DMF solution and 3.2mL of DIC, and let it react for 15min. Then add the solution in the 100mL beaker to the above 250mL solid-phase synthesis reactor, stir and react for 1.5h, and the reaction is complete; wash the resin with DMF solution 3 times, 75mL each time; after washing, proceed to the next step of reaction; add 75mL of 20% Pip / DMF (v / v) solution, stir and react for 30min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 65mL each time, and drain for use.

[0052] Take 21mmol Fmoc-Thr(tBu)-OH and 21mmol HOBt in a 100mL beaker, cool to 5℃, add 50mL of DMF solution and 3.2mL of DIC, and let it react for 15min. Then add the solution in the 100mL beaker to the above 250mL solid-phase synthesis reactor, stir and react for 1.5h, and the reaction is completed; wash the resin with DMF solution 3 times, 75mL each time; after washing, proceed to the next step of reaction; add 75mL of 20% Pip / DMF (v / v) solution, stir and react for 30min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 65mL each time, and drain for use.

[0053] 21 mmol Fmoc-Lys(Boc)-OH and 21 mmol HOBt were placed in a 100 mL beaker and cooled to 2-8°C. 50 mL of DMF solution and 3.2 mL of DIC were added and the mixture was allowed to react for 15 min. The solution in the 100 mL beaker was added to the above 250 mL solid phase synthesis reactor and stirred for 1.5 h. The reaction was completed. The resin was washed with DMF solution three times, 75 mL each time. After washing, the next step was carried out. 75 mL of 20% Pip / DMF (v / v) solution was added and stirred for 30 min. The deprotection solution was removed by filtration. The resin was then washed with DMF solution six times, 65 mL each time, methanol twice, 75 mL each time, DCM solution twice, and methanol twice, 75 mL each time. The resin was then dried under vacuum to obtain H-Lys(Boc)-Thr(tBu)-Thr(tBu)-Lys(Boc)-Ser(tBu)-CTC-resin. 4.14 g of the above peptide resin was added with 40 mL of cutting solution TFA / Tis / H2O (the volume ratio of TFA, Tis and H2O was 90:5:5), and the mixture was stirred at 30°C for 2.5 h. The resin was removed by filtration to obtain a filtrate. The filtrate was dried to obtain H-Lys-Thr-Thr-Lys-Ser-OH, and its mass spectrometry and HPLC characterization results are shown in the following table. Figure 5 and Figure 6 shown.

[0054] The preparation method of hyaluronic acid-modified cosmetic peptide comprises the following steps: Weigh 0.5 g of H-Lys-Thr-Thr-Lys-Ser-OH.3TFA, add 5 mL of DMSO and stir to dissolve, add 82 μL of TFA, stir for 0.5 h, then add 0.641 g of DIEA, weigh 1.71 g of hyaluronic acid, control the temperature in a water bath at 45°C, stir and react overnight, take a sample and detect by LC-MS, if the reaction is basically complete, add 4 mL of acetic acid, control the temperature in a water bath at 35°C, and carry out the rearrangement reaction for 3 h, take a sample and detect by LC-MS, if the reaction is basically complete; then perform reverse chromatography purification, purification conditions: Dissolution: dilute 0.5 g of crude product with 100 mL of H2O; Packing: 21.2*250mm, 10-120, C18; Flow rate: 10mL / min; Wavelength: 220nm; Mobile phase: A: 1% HAc; B: ACN; Balance: A:B=100:0, balance 10 min, flow rate: 10 mL / min; Sample loading: flow rate: 10 mL / min; Elution: 0-10% B 60 min; Column cleaning: 80% ACN cleaning to baseline equilibrium; Qualified products were collected to obtain hyaluronic acid modified beauty peptides, which were characterized by high performance liquid chromatography. Figure 7 As shown; Among them, samples at 5.801min, 6.102min, and 6.558min were collected for mass spectrometry characterization, and the results are as follows Figure 8 、 Figure 9 、 Figure 10 shown.

[0055] Example 3: A method for synthesizing H-Lys-Leu-Ala-Lys-Lys-NH2, comprising: Place 10 mmol of AM resin in a 250 mL solid-phase synthesis reactor, add 70 mL of 20% Pip / DMF (v / v) solution, stir and react for 30 min, filter to remove the deprotection solution, then wash with DMF solution 6 times, 70 mL each time, and drain to dryness for use.

[0056] Take 20mmol Fmoc-Linker and 20mmol HOBt in a 100mL beaker, cool to 5℃, add 35mL DMF solution and 3.1mL DIC, let it react for 15min, and add the solution in the 100mL beaker to the above 250mL solid-phase synthesis reactor. Stir and react for 1.5h. The reaction is complete; wash the resin with DMF solution 3 times, 70mL each time; after washing, proceed to the next step; add 70mL of 20% Pip / DMF (v / v) solution, stir and react for 30min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 70mL each time, and drain and set aside.

[0057] Take 20mmol Fmoc-Lys(Boc)-OH and 20mmol HOBt in a 100mL beaker, cool to 5℃, add 35mL DMF solution and 3.1mL DIC, let it stand for 15min, and add the solution in the 100mL beaker to the above 250mL solid phase synthesis reactor. Stir and react for 1.5h. The reaction is complete; wash the resin with DMF solution 3 times, 70mL each time; after washing, proceed to the next step; add 70mL of 20% Pip / DMF (v / v) solution, stir and react for 30min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 70mL each time, and drain for use.

[0058] Take 20mmol Fmoc-Lys(Boc)-OH and 20mmol HOBt in a 100mL beaker, cool to 5℃, add 35mL DMF solution and 3.1mL DIC, let it stand for 15min, and add the solution in the 100mL beaker to the above 250mL solid phase synthesis reactor. Stir and react for 1.5h. The reaction is complete; wash the resin with DMF solution 3 times, 70mL each time; after washing, proceed to the next step; add 70mL of 20% Pip / DMF (v / v) solution, stir and react for 30min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 70mL each time, and drain for use.

[0059] Take 20mmol Fmoc-Ala-OH and 20mmol HOBt in a 100mL beaker, cool to 5℃, add 35mL DMF solution and 3.1mL DIC and let it react for 15min. Then add the solution in the 100mL beaker to the above 250mL solid-phase synthesis reactor and stir to react for 1.5h. The reaction is completed. Wash the resin with DMF solution 3 times, 70mL each time. After washing, proceed to the next step of reaction. Add 70mL of 20% Pip / DMF (v / v) solution, stir to react for 30min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 70mL each time, and drain for use.

[0060] Take 20mmol Fmoc-Leu-OH and 20mmol HOBt in a 100mL beaker, cool to 5℃, add 35mL DMF solution and 3.1mL DIC, let it stand for 15min, and add the solution in the 100mL beaker to the above 250mL solid-phase synthesis reactor. Stir and react for 1.5h. The reaction is complete; wash the resin with DMF solution 3 times, 70mL each time; after washing, proceed to the next step; add 70mL of 20% Pip / DMF (v / v) solution, stir and react for 30min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 40mL each time, and drain for use.

[0061] 20 mmol of Fmoc-Lys(Boc)-OH and 20 mmol of HOBt were placed in a 100 mL beaker and cooled to 5°C. 35 mL of DMF solution and 3.1 mL of DIC were added and the mixture was allowed to react for 15 minutes. The solution in the 100 mL beaker was then added to the 250 mL solid-phase synthesis reactor described above and stirred for 1.5 hours. The reaction was completed. The resin was washed with DMF solution three times, 70 mL each time. After washing, the resin was processed to the next step. 70 mL of 20% Pip / DMF (v / v) solution was added and stirred for 30 minutes. The deprotection solution was removed by filtration and then washed with DMF solution six times, 40 mL each time, and dried. The resin was then washed with methanol twice, 70 mL each time, DCM twice, and methanol twice, 70 mL each time. The resin was dried under vacuum to obtain the peptide resin H-Lys(Boc)-Leu-Ala-Lys(Boc)-Lys(Boc)-Linker-AM resin. To the above peptide resin, 100 mL of cutting solution TFA / Tis / H2O (the volume ratio of TFA, Tis and H2O is 90:5:5) was added and cut for 2.5 hours. The cutting solution was added to 1000 mL of ether (5°C) solution to precipitate a white solid, which was centrifuged and vacuum dried to obtain H-Lys-Leu-Ala-Lys-Lys-NH2. Its mass spectrum and LC-MS characterization results are shown in Figure 2. Figure 11 and Figure 12 shown.

[0062] The preparation method of hyaluronic acid-modified cosmetic peptide comprises the following steps: Weigh 0.5 g of H-Lys-Leu-Ala-Lys-Lys-NH2.4TFA, add 10 mL of DMSO and stir to dissolve, add 107 μL of TFA, stir at room temperature for 0.5 h, add 0.557 g of DIEA, then add 1.49 g of hyaluronic acid, control the temperature in a water bath at 45°C, stir and react overnight, take a sample and detect by LC-MS, if the reaction is basically complete, add 4 mL of acetic acid, control the temperature in a water bath at 35°C, and carry out the rearrangement reaction for 3 h, take a sample and detect by LC-MS, if the reaction is basically complete; then perform reverse chromatography purification, purification conditions: Dissolution: dilute 0.5 g of crude product with 100 mL of H2O; Packing: 21.2*250mm, 10-120, C18; Flow rate: 10mL / min; Wavelength: 220nm; Mobile phase: A: 1% HAc; B: ACN; Balance: A:B=100:0, balance 10 min, flow rate: 10 mL / min; Sample loading: flow rate: 10 mL / min; Elution: 0-10% B 60 min; Column cleaning: 80% ACN cleaning to baseline equilibrium; Qualified products were collected to obtain hyaluronic acid modified beauty peptides, and their mass spectrometry and HPLC characterization results were as follows: Figure 13 and Figure 14 shown.

[0063] Example 4: A method for synthesizing H-Glu-Glu-Met-Gln-Arg-Arg-NH2, comprising: Place 5 mmol of AM resin in a 100 mL solid-phase synthesis reactor, add 30 mL of 20% Pip / DMF (v / v) solution, stir and react for 30 min, filter to remove the deprotection solution, then wash with DMF solution 6 times, 30 mL each time, and drain to dryness for later use; Take 10mmol Fmoc-Linker and 10mmol HOBt in a 100mL beaker, cool to 5℃, add 15mL of DMF solution and 1.5mL of DIC and let it react for 15min. Then add the solution in the 100mL beaker to the above 100mL solid-phase synthesis reactor and stir to react for 1.5h. The reaction is complete. Wash the resin with DMF solution 3 times, 20mL each time. After washing, proceed to the next step of reaction. Add 30mL of 20% Pip / DMF (v / v) solution, stir to react for 30min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 30mL each time, and drain for use.

[0064] Place 10 mmol of Fmoc-Arg(Pbf)-OH and 10 mmol of HOBt in a 100 mL beaker, cool to 5°C, add 15 mL of DMF solution and 1.5 mL of DIC, and let the mixture react for 15 minutes. The mixture was then added to the 100 mL solid-phase synthesis reactor described above and stirred for 1.5 hours. The reaction was complete. The resin was washed three times with 20 mL of DMF solution. After washing, the resin was moved to the next step: 30 mL of a 20% Pip / DMF (v / v) solution was added and stirred for 30 minutes. The deprotection solution was removed by filtration, and the resin was then washed six times with 30 mL of DMF solution. The resin was then dried and used.

[0065] Take 10mmol Fmoc-Arg(Pbf)-OH and 10mmol HOBt in a 100mL beaker, cool to 5℃, add 15mL of DMF solution and 1.5mL of DIC and let it react for 15min. Then add the solution in the 100mL beaker to the above 100mL solid phase synthesis reactor and stir to react for 1.5h. The reaction is completed. Wash the resin with DMF solution 3 times, 20mL each time. After washing, proceed to the next step of reaction. Add 30mL of 20% Pip / DMF (v / v) solution, stir to react for 30min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 30mL each time, and dry for use.

[0066] Take 10mmol Fmoc-Gln (Trt) -OH and 10mmol HOBt in a 100mL beaker, cool to 5℃, add 15mL DMF solution and 1.5mL DIC and let it react for 15min. Then add the solution in the 100mL beaker to the above 100mL solid phase synthesis reactor and stir to react for 1.5h. The reaction is completed. Wash the resin with DMF solution 3 times, 20mL each time. After washing, proceed to the next step of reaction. Add 30mL of 20% Pip / DMF (v / v) solution, stir to react for 30min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 30mL each time, and dry for use.

[0067] Take 10mmol Fmoc-Met-OH and 10mmol HOBt in a 100mL beaker, cool to 5℃, add 15mL of DMF solution and 1.5mL of DIC, and let it react for 15min. Then add the solution in the 100mL beaker to the above 100mL solid-phase synthesis reactor, stir and react for 1.5h, and the reaction is completed; wash the resin with DMF solution 3 times, 20mL each time; after washing, proceed to the next step of reaction; add 40mL of 20% Pip / DMF (v / v) solution, stir and react for 30min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 40mL each time, and drain and set aside.

[0068] Take 10mmol Fmoc-Glu(otBu)-OH and 10mmol HOBt in a 100mL beaker, cool to 5℃, add 15mL of DMF solution and 1.5mL of DIC and let it react for 15min. Then add the solution in the 100mL beaker to the above 100mL solid phase synthesis reactor and stir to react for 1.5h. The reaction is completed. Wash the resin with DMF solution 3 times, 20mL each time. After washing, proceed to the next step of reaction. Add 40mL of 20% Pip / DMF (v / v) solution, stir to react for 30min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 40mL each time, and drain for use.

[0069] Take 15mmo Fmoc-Glu(otBu)-OH, 1mmol HOBt was placed in a 100 mL beaker and cooled to 5°C. 15 mL of DMF solution and 2.3 mL of DMF solution were added and allowed to react for 15 min. The solution in the 100 mL beaker was added to the above 100 mL solid phase synthesis reactor and stirred for 1.5 h. The reaction was completed. The resin was washed with DMF solution 3 times, 30 mL each time. After washing, the next step was carried out. 40 mL of 20% Pip / DMF (v / v) solution was added and stirred for 30 min. The deprotection solution was removed by filtration and then washed with DMF solution 6 times, 40 mL each time, and dried for use. Then, methanol was added for washing 2 times, 40 mL each time, DCM solution was added 2 times, 40 mL each time, and methanol was added 2 times, 40 mL each time, and vacuum dried to obtain the peptide resin of H-Glu(otBu)-Glu(otBu)-Met-Gln(Trt)-Arg(Pbf)-Arg(Pbf)-Linker-AM resin. The peptide resin was cleaved with 100 mL of TFA / thioanisole / phenol / H2O / EDT (the mass ratio of TFA, thioanisole, phenol, H2O and EDT was 87.5:5:2.5:2.5:2.5) for 2.5 h. The cleavage solution was added to 1000 mL of ether (5°C) to precipitate a white solid, which was centrifuged and dried in vacuo to obtain H-Glu-Glu-Met-Gln-Arg-Arg-NH2. Its mass spectrometry and HPLC characterization results are shown in Figure 2. Figure 15 and Figure 16 shown.

[0070] A synthetic route for a hyaluronic acid-modified beauty peptide is: ; Wherein, -COOX is -COONa; The preparation method of hyaluronic acid-modified cosmetic peptide comprises the following steps: Weigh 1g of H-Glu-Glu-Met-Gln-Arg-Arg-NH2.3TFA, add 10mL of DMSO and stir to dissolve, add 889mg of DIEA, then add 3.67g of hyaluronic acid, control the temperature in a water bath at 45°C, stir and react overnight, take a sample and detect by LC-MS, if the reaction is basically complete, add 8mL of acetic acid, control the temperature in a water bath at 35°C, and carry out the rearrangement reaction for 3h, take a sample and detect by LC-MS, if the reaction is basically complete; then perform reverse phase chromatography purification, purification conditions: Dissolution: dilute 1 g of crude product with 200 mL of H2O; Filler: 50DAC, C18; Flow rate: 60 mL / min; Wavelength: 220 nm; Mobile phase: A: 1% HAc; B: ACN; Balance: A:B=100:0, balance 10 min, flow rate: 60 mL / min; Sample loading: flow rate: 60 mL / min; Elution: 0-20%B 60 min; Column cleaning: 80% ACN cleaning to baseline equilibrium; Qualified products were collected to obtain three structures of hyaluronic acid-modified beauty peptides, as shown below: D2, its mass spectrometry and HPLC characterization results are shown in Figure 17 and Figure 18 As shown; D4, its mass spectrometry and HPLC characterization results are shown in Figure 19 and Figure 20 As shown; D6, its mass spectrometry and HPLC characterization results are shown in Figure 21 and Figure 22 shown.

[0071] Example 5: The preparation method of hyaluronic acid-modified cosmetic peptide comprises the following steps: Weigh 0.5g H-Gly-His-Lys-OH.2AcOH, add 10mL DMSO and stir for 5min to dissolve, add 286μLTFA, stir for 1min until the solution becomes clear, stir for another 0.5h, add 3g hyaluronic acid, stir to dissolve, add 1.11g DIEA, stir for 1min, control the temperature in a water bath at 45℃, stir and react overnight, take a sample and detect LC-MS, the reaction is basically complete, add 4mL acetic acid, control the temperature in a water bath at 35℃, and carry out the rearrangement reaction for 3h, take a sample and detect LC-MS, the reaction is basically complete; then perform reverse chromatography purification, purification conditions: Dissolution: dilute 0.5 g of crude product with 100 mL of H2O; Packing: 21.2*250mm, 10-120, C18; Flow rate: 10mL / min; Wavelength: 220nm; Mobile phase: A: 1% HAc; B: ACN; Balance: A:B=100:0, balance 10 min, flow rate: 10 mL / min; Sample loading: flow rate: 10 mL / min; Elution: 0-20%B 60 min; Column cleaning: 80% ACN cleaning to baseline equilibrium; Qualified products were collected to obtain hyaluronic acid modified beauty peptides, and their mass spectrometry and HPLC test results were as follows: Figure 23 and Figure 24 shown.

[0072] Example 6: A method for synthesizing H-Gly-Pro-Gln-Gly-Pro-Gln-OH, comprising: Place 10 mmol of AM resin in a 100 mL solid-phase synthesis reactor, add 30 mL of 20% Pip / DMF (v / v) solution, stir and react for 30 min, filter to remove the deprotection solution, then wash with DMF solution 6 times, 30 mL each time, and drain to dryness for use.

[0073] Take 20mmol Fmoc-Linker and 20mmol HOBt in a 100mL beaker, cool to 5℃, add 15mL DMF solution and 3.1mL DIC and let it react for 15min. Then add the solution in the 100mL beaker to the above 100mL solid-phase synthesis reactor and stir to react for 1.5h. The reaction is complete. Wash the resin with DMF solution 3 times, 20mL each time. After washing, proceed to the next step of reaction. Add 30mL of 20% Pip / DMF (v / v) solution, stir to react for 30min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 30mL each time, and drain for use.

[0074] Take 20mmol Fmoc-Glu-OtBu and 20mmol HOBt in a 100mL beaker, cool to 5℃, add 15mL DMF solution and 4.6mL DIC, let it react for 15min, and add the solution in the 100mL beaker to a 100mL solid-phase synthesis reactor. Stir and react for 1.5h until the reaction is complete. Wash the resin with DMF solution 3 times, 20mL each time. After washing, proceed to the next step. Add 30mL of 20% Pip / DMF (v / v) solution, stir and react for 30min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 30mL each time, and dry it for use.

[0075] Take 20mmol Fmoc-Pro-OH and 20mmol HOBt in a 100mL beaker, cool to 5℃, add 15mL of DMF solution and 4.6mL of DIC, and let it react for 15min. Then add the solution in the 100mL beaker to the above 100mL solid-phase synthesis reactor, stir and react for 1.5h, and the reaction is complete; wash the resin with DMF solution 3 times, 20mL each time; after washing, proceed to the next step of reaction; add 30mL of 20% Pip / DMF (v / v) solution, stir and react for 30min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 30mL each time, and dry it for use.

[0076] Take 20mmo Fmoc-Gly-OH and 20mmo HOBt in a 100mL beaker, cool to 5℃, add 15mL of DMF solution and 4.6mL of DIC, and let it react for 15min. Then add the solution in the 100mL beaker to the above 100mL solid-phase synthesis reactor, stir and react for 1.5h, and the reaction is complete; wash the resin with DMF solution 3 times, 20mL each time; after washing, proceed to the next step of reaction; add 30mL of 20% Pip / DMF (v / v) solution, stir and react for 30min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 30mL each time, and drain for use.

[0077] Take 20mmol Fmoc-Gln(Trt)-OH and 20mmol HOBt in a 100mL beaker, cool to 5℃, add 15mL of DMF solution and 4.6mL of DIC, and let it react for 15min. Then add the solution in the 100mL beaker to the above 100mL solid-phase synthesis reactor, stir and react for 1.5h, and the reaction is completed; wash the resin with DMF solution 3 times, 20mL each time; after washing, proceed to the next step of reaction; add 40mL of 20% Pip / DMF (v / v) solution, stir and react for 30min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 40mL each time, and dry it for use.

[0078] Take 20mmol Fmoc-Pro-OH and 20mmol HOBt in a 100mL beaker, cool to 5℃, add 15mL of DMF solution and 4.6mL of DIC, and let it react for 15min. Then add the solution in the 100mL beaker to the above 100mL solid-phase synthesis reactor, stir and react for 1.5h, and the reaction is complete; wash the resin with DMF solution 3 times, 20mL each time; after washing, proceed to the next step of reaction; add 30mL of 20% Pip / DMF (v / v) solution, stir and react for 30min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 40mL each time, and drain and set aside.

[0079] 20 mmol Fmoc-Gly-OH and 20 mmol HOBt were placed in a 100 mL beaker and cooled to 5°C. 15 mL of DMF solution and 4.6 mL of DIC were added and the mixture was allowed to react for 15 minutes. The solution in the 100 mL beaker was then added to the 100 mL solid-phase synthesis reactor described above and stirred for 1.5 hours. The reaction was completed. The resin was washed three times with 30 mL of DMF solution. After washing, the resin was subjected to the next step. 30 mL of 20% Pip / DMF (v / v) solution was added and stirred for 30 minutes. The deprotection solution was removed by filtration and then washed six times with 40 mL of DMF solution. The resin was then dried and used. The resin was then washed twice with 40 mL of methanol, twice with 40 mL of DCM solution, and twice with 40 mL of methanol. Drying under vacuum gave H-BB1-BB2-BB3-BB4-BB5-BB6-Linker-AM resin, wherein AA1 is Gly; AA2 is Pro; AA3 is Gln; AA4 is Gly; AA5 is Pro; and AA6 is Gln.

[0080] 8.3 g of the above peptide resin was cleaved with 50 mL of cleavage solution (TFA, Tis, and H2O in a volume ratio of 90:5:5) for 2.5 h. The cleavage solution was added to 500 mL of ether (5°C) to precipitate a white solid, which was centrifuged and dried in vacuo to obtain H-Gly-Pro-Gln-Gly-Pro-Gln-OH. Its mass spectrometry and HPLC characterization results are shown in Figure 2. Figure 25 and Figure 26 shown.

[0081] The preparation method of hyaluronic acid-modified cosmetic peptide comprises the following steps: Weigh 0.5g of H-Gly-Pro-Gln-Gly-Pro-Gln-OH. TFA, add 5mL of DMSO and stir to dissolve, then add 0.834g of DIEA and 2.23g of hyaluronic acid. Stir and react overnight in a water bath at 45°C. Samples were taken for LC-MS analysis, indicating that the reaction was essentially complete. Add 4mL of acetic acid, control the temperature in a water bath at 35°C, and allow the rearrangement reaction to proceed for 3h. Samples were taken for LC-MS analysis, indicating that the reaction was essentially complete. Then, perform reversed-phase chromatography purification under the following purification conditions: Dissolution: dilute 0.5 g of crude product with 100 mL of H2O; Packing: 21.2*250mm, 10-120, C18; Flow rate: 10mL / min; Wavelength: 220nm; Mobile phase: A: 1% HAc; B: ACN; Balance: A:B=100:0, balance 10 min, flow rate: 10 mL / min; Sample loading: flow rate: 10 mL / min; Elution: 0-20%B 60 min; Column cleaning: 80% ACN cleaning to baseline equilibrium; Qualified products were collected to obtain hyaluronic acid modified beauty peptides, and their mass spectrometry and HPLC characterization results were as follows: Figure 27 and Figure 28 shown.

[0082] Example 7: A method for synthesizing H-Met-Pro-{D-Phe}-Arg-{D-Trp}-Phe-Lys-Pro-Val-NH2 comprises: Place 5 mmol AM resin in a 100 mL solid-phase synthesis reactor, add 30 mL of 20% Pip / DMF (v / v) solution, stir and react for 30 min, filter to remove the deprotection solution, then wash with DMF solution 6 times, 30 mL each time, and drain to set aside.

[0083] Place 10 mmol of Fmoc-Linker and 10 mmol of HOBt in a 100 mL beaker, cool to 5°C, add 15 mL of DMF solution and 1.5 mL of DIC, and let the mixture react for 15 minutes. The solution in the 100 mL beaker was then added to the 100 mL solid-phase synthesis reactor described above, and the reaction was stirred for 1.5 hours until the reaction was complete. The resin was washed three times with 20 mL of DMF solution. After washing, the resin was moved to the next step: 30 mL of 20% Pip / DMF (v / v) solution was added, and the mixture was stirred for 30 minutes. The deprotection solution was removed by filtration, and the resin was then washed six times with 30 mL of DMF solution, dried, and set aside.

[0084] Take 15mmo Fmoc-Val-OH and 15mmol HOBt in a 100mL beaker, cool to 5℃, add 15mL of DMF solution and 2.3mL of DIC and let it react for 15min. Then add the solution in the 100mL beaker to the above 100mL solid-phase synthesis reactor and stir to react for 1.5h. The reaction is complete. Wash the resin with DMF solution 3 times, 20mL each time. After washing, proceed to the next step of reaction. Add 30mL of 20% Pip / DMF (v / v) solution, stir to react for 30min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 30mL each time, and drain for use.

[0085] Take 15mmol Fmoc-Pro-OH and 15mmol HOBt in a 100mL beaker, cool to 5℃, add 15mL of DMF solution and 2.3mL of DIC, and let it react for 15min. Then add the solution in the 100mL beaker to the above 100mL solid-phase synthesis reactor, stir and react for 1.5h, and the reaction is completed; wash the resin with DMF solution 3 times, 20mL each time; after washing, proceed to the next step of reaction; add 30mL of 20% Pip / DMF (v / v) solution, stir and react for 30min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 30mL each time, and dry it for use.

[0086] Take 15mmol Fmoc-Lys(Boc)-OH and 15mmol HOBt in a 100mL beaker, cool to 5℃, add 15mL of DMF solution and 2.3mL of DIC and let it react for 15min. Then add the solution in the 100mL beaker to the above 100mL solid phase synthesis reactor and stir to react for 1.5h. The reaction is completed. Wash the resin with DMF solution 3 times, 20mL each time. After washing, proceed to the next step of reaction. Add 30mL of 20% Pip / DMF (v / v) solution, stir to react for 30min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 30mL each time, and drain for use.

[0087] Take 15mmol Fmoc-Phe-OH and 15mmol HOBt in a 100mL beaker, cool to 5℃, add 15mL of DMF solution and 2.3mL of DIC and let it react for 15min. Then add the solution in the 100mL beaker to the above 100mL solid-phase synthesis reactor and stir to react for 1.5h. The reaction is completed. Wash the resin with DMF solution 3 times, 20mL each time. After washing, proceed to the next step of reaction. Add 40mL of 20% Pip / DMF (v / v) solution, stir to react for 30min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 40mL each time, and drain for use.

[0088] Take 10mmol Fmoc-D-Trp(Boc)-OH and 10mmol HOBt in a 100mL beaker, cool to 5℃, add 15mL of DMF solution and 1.5mL of DIC and let it react for 15min. Then add the solution in the 100mL beaker to the above 100mL solid phase synthesis reactor and stir to react for 1.5h. The reaction is completed. Wash the resin with DMF solution 3 times, 20mL each time. After washing, proceed to the next step of reaction. Add 40mL of 20% Pip / DMF (v / v) solution, stir to react for 30min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 40mL each time, and drain for use.

[0089] Place 15 mmol of Fmoc-Arg(Pbf)-OH and 15 mmol of HOBt in a 100 mL beaker, cool to 5°C, add 15 mL of DMF solution and 2.3 mL of DIC, and let the mixture react for 15 minutes. The mixture was then added to the 100 mL solid-phase synthesis reactor described above and stirred for 1.5 hours. The reaction was complete. The resin was washed three times with 30 mL of DMF solution. After washing, the resin was moved to the next step: 40 mL of a 20% Pip / DMF (v / v) solution was added and stirred for 30 minutes. The deprotection solution was removed by filtration, and the resin was then washed six times with 40 mL of DMF solution. The resin was then dried and used.

[0090] Take 10mmol Fmoc-D-Phe-OH and 10mmol HOBt in a 100mL beaker, cool to 5℃, add 15mL of DMF solution and 1.5mL of DIC, and let it react for 15min. Then add the solution in the 100mL beaker to the above 100mL solid-phase synthesis reactor, stir and react for 1.5h, and the reaction is complete; wash the resin with DMF solution 3 times, 30mL each time; after washing, proceed to the next step of reaction; add 40mL of 20% Pip / DMF (v / v) solution, stir and react for 30min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 40mL each time, and dry it for use.

[0091] Take 15mmol Fmoc-Pro-OH and 15mmol HOBt in a 100mL beaker, cool to 5℃, add 15mL of DMF solution and 2.3mL of DIC, and let it react for 15min. Then add the solution in the 100mL beaker to the above 100mL solid-phase synthesis reactor, stir and react for 1.5h, and the reaction is completed; wash the resin with DMF solution 3 times, 30mL each time; after washing, proceed to the next step of reaction; add 40mL of 20% Pip / DMF (v / v) solution, stir and react for 30min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 40mL each time, and drain for use.

[0092] 15 mmol Fmoc-Met-OH and 15 mmol HOBt were placed in a 100 mL beaker and cooled to 5°C. 15 mL of DMF solution and 2.3 mL of DIC were added and the mixture was allowed to react for 15 minutes. The solution in the 100 mL beaker was then added to the 100 mL solid-phase synthesis reactor described above and stirred for 1.5 hours. The resin was then washed with DMF solution three times, 30 mL each time. After washing, the resin was subjected to the next step. 40 mL of 20% Pip / DMF (v / v) solution was added and stirred for 30 minutes. The deprotection solution was removed by filtration and then washed with DMF solution six times, 40 mL each time, and dried. The resin was then washed with methanol twice, 40 mL each time, DCM twice, and methanol twice, 40 mL each time. The resin was then dried under vacuum to obtain the peptide resin H-Met-Pro-{D-Phe}-Arg-{D-Trp}-Phe-Lys-Pro-Val-Linker-AM resin.

[0093] 5.7 g of the above peptide resin was cleaved with 50 mL of a cleavage solution of TFA / thioanisole / EDT / phenol / H2O (TFA:thioanisole:EDT:phenol:H2O mass ratio of 87.5:5:2.5:2.5:2.5) for 2.5 h. The cleavage solution was added to 500 mL of ether (5°C) to precipitate a white solid, which was centrifuged and dried in vacuo to obtain H-Met-Pro-{D-Phe}-Arg-{D-Trp}-Phe-Lys-Pro-Val-NH2. Its mass spectrum and high performance liquid chromatography were as shown in FIG. Figure 29 and Figure 30 shown.

[0094] The preparation method of hyaluronic acid-modified cosmetic peptide comprises the following steps: Weigh 0.5 g of H-Met-Pro-{D-Phe}-Arg-{D-Trp}-Phe-Lys-Pro-Val-NH2.3TFA, add 5 mL of DMSO and stir to dissolve, add 0.375 g of DIEA, then add 1 g of hyaluronic acid, control the temperature in a water bath at 45°C, stir and react overnight, take a sample and detect by LC-MS, if the reaction is basically complete, add 4 mL of acetic acid, control the temperature in a water bath at 35°C, and carry out the rearrangement reaction for 3 hours, take a sample and detect by LC-MS, if the reaction is basically complete; then perform reverse chromatography purification, purification conditions: Dissolution: dilute 0.5 g of crude product with 100 mL of H2O; Packing: 21.2*250mm, 10-120, C18; Flow rate: 10mL / min; Wavelength: 220nm; Mobile phase: A: 1% HAc; B: ACN; Balance: A:B=95:5, balance 10 min, flow rate: 10 mL / min; Sample loading: flow rate: 10 mL / min; Elution: 15-35%B 60 min; Column cleaning: 80% ACN cleaning to baseline equilibrium; Qualified products were collected to obtain hyaluronic acid modified beauty peptides, and their mass spectrometry and HPLC characterization results were as follows: Figure 31 and Figure 32 shown.

[0095] Example 8: A method for synthesizing H-Phe-Val-Ala-Pro-Phe-Pro-OH, comprising: Place 12.5 mmol of CTC resin in a 125 mL solid-phase synthesis reactor, add 25 mmol of amino acid Fmoc-Pro-OH, add 100 mL of dichloromethane, then add 10.9 mL of DIEA, react at 25 ° C for 3 h, add 15 mL of methanol, and react for 5 min; filter, wash the resin with dichloromethane twice, 100 mL each time, methanol twice, 100 mL each time, and DMF twice, 100 mL each time; add 100 mL of 20% Pip / DMF (v / v) solution, stir and react for 30 min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 20 mL each time, and dry for use.

[0096] Take 20mmol Fmoc-Phe-OH and 20mmol HOBt in a 50mL beaker, cool to 5℃, add 30mL DMF solution and 3.1mL DIC, let it stand for 15min, and add the solution in the 100mL beaker to the above 250mL solid-phase synthesis reactor. Stir and react for 1.5h until the reaction is complete. Wash the resin with DMF solution 3 times, 100mL each time. After washing, proceed to the next step of reaction. Add 100mL of 20% Pip / DMF (v / v) solution, stir and react for 30min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 20mL each time, and drain and set aside.

[0097] Take 20mmol Fmoc-Pro-OH and 20mmol HOBt in a 50mL beaker, cool to 5℃, add 30mL DMF solution and 3.1mL DIC, let it stand for 15min, and add the solution in the 100mL beaker to the above 250mL solid-phase synthesis reactor. Stir and react for 1.5h. The reaction is complete; wash the resin with DMF solution 3 times, 100mL each time; after washing, proceed to the next step; add 100mL of 20% Pip / DMF (v / v) solution, stir and react for 30min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 20mL each time, and drain for use.

[0098] Take 20mmo Fmoc-Ala-OH and 20mmol HOBt in a 50mL beaker, cool to 5℃, add 8mL of DMF solution and 3.1mL of DIC and let it react for 15min. Then add the solution in the 100mL beaker to the above 250mL solid-phase synthesis reactor and stir to react for 1.5h. The reaction is complete. Wash the resin with DMF solution 3 times, 100mL each time. After washing, proceed to the next step of reaction. Add 100mL of 20% Pip / DMF (v / v) solution, stir to react for 30min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 20mL each time, and drain for use.

[0099] Take 20mmol Fmoc-Val-OH and 20mmol HOBt in a 50mL beaker, cool to 5℃, add 8mL of DMF solution and 20mmol DIC, let it stand for 5min, and add the solution in the 100mL beaker to the above 250mL solid-phase synthesis reactor. Stir and react for 1.5h until the reaction is complete. Wash the resin with DMF solution 3 times, 100mL each time. After washing, proceed to the next step of reaction. Add 100mL of 20% Pip / DMF (v / v) solution, stir and react for 30min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 20mL each time, and drain and set aside.

[0100] Take 20mmol Fmoc-Phe-OH and 20mmol HOBt in a 50mL beaker, cool to 5℃, add 30mL DMF solution and 3.1mL DIC, let it stand for 15min, and add the solution in the 100mL beaker to the above 250mL solid phase synthesis reactor, stir and react for 1.5h, and the reaction is complete; wash the resin with DMF solution 3 times, 100mL each time; after washing, proceed to the next step; add 100mL of 20% Pip / DMF (v / v) solution, stir and react for 30min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 20mL each time, then wash with methanol 2 times, 100mL each time, wash with DCM solution 2 times, 100mL each time, wash with methanol 2 times, 100mL each time, and dry in vacuum to obtain H-Phe-Val-Ala-Pro-Phe-Pro-CTC-resin; 2 g of the above peptide resin was cleaved with 20 mL of TFA / DCM (volume ratio of TFA to DCM was 2:98) at 30°C for 1 hour, and the cleavage solution was dried to obtain H-Phe-Val-Ala-Pro-Phe-Pro-OH. The mass spectrum and LC-MS characterization results are shown in Figure 2. Figure 33 and Figure 34 As shown. A synthetic route of a hyaluronic acid modified beauty peptide is: ; Wherein, -COOX is -COONa; The preparation method of hyaluronic acid-modified cosmetic peptide comprises the following steps: Weigh 0.56g H-Phe-Val-Ala-Pro-Phe-Pro-OH.TFA, add 6mL DMSO and stir to dissolve, add 0.75g DIEA, then add 2.2g hyaluronic acid, control the temperature in a water bath at 45°C, stir and react overnight, take a sample and detect by LC-MS, the reaction is basically complete, add 4mL acetic acid, control the temperature in a water bath at 35°C, and carry out the rearrangement reaction for 3h, take a sample and detect by LC-MS, the reaction is basically complete; then perform reverse chromatography purification, purification conditions: Dissolution: dilute 0.5 g of crude product with 100 mL of H2O; Packing: 21.2*250mm, 10-120, C18; Flow rate: 10mL / min; Wavelength: 220nm; Mobile phase: A: 1% HAc; B: ACN; Balance: A:B=95:5, balance 10 min, flow rate: 10 mL / min; Sample loading: flow rate: 10 mL / min; Elution: 12-32%B 60 min; Column cleaning: 80% ACN cleaning to baseline equilibrium; Qualified products were collected to obtain three types of hyaluronic acid-modified beauty peptides, namely: Hyaluronic acid modified beauty peptide H2, in which n=0 in its structure, i.e. y=1 in the raw material hyaluronic acid structure, and the product prepared with the beauty peptide, its mass spectrometry and HPLC characterization results are as follows Figure 35 and Figure 36 As shown; Hyaluronic acid modified beauty peptide H4, in which n=1 in its structure, i.e. y=2 in the raw material hyaluronic acid structure, and the product prepared with the beauty peptide, its mass spectrometry and HPLC characterization results are as follows Figure 37 and Figure 38 As shown; Hyaluronic acid modified beauty peptide H6, in which n=2 in its structure, i.e. y=3 in the raw material hyaluronic acid structure, and the product prepared with the beauty peptide, its mass spectrometry and HPLC characterization results are as follows Figure 39 and Figure 40 shown.

[0101] Example 9: A method for synthesizing H-His-D-Phe-Arg-NH2, comprising: Place 10 mmol of AM resin in a 250 mL solid-phase synthesis reactor, add 70 mL of 20% Pip / DMF (v / v) solution, stir and react for 30 min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 70 mL each time, and drain to use.

[0102] Take 20mmol Fmoc-Linker and 20mmol HOBt in a 100mL beaker, cool to 5℃, add 35mL DMF solution and 3.1mL DIC, let it react for 15min, and add the solution in the 100mL beaker to the above 250mL solid-phase synthesis reactor. Stir and react for 1.5h. The reaction is complete; wash the resin with DMF solution 3 times, 70mL each time; after washing, proceed to the next step; add 70mL of 20% Pip / DMF (v / v) solution, stir and react for 30min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 70mL each time, and drain and set aside.

[0103] Take 20mmol Fmoc-Arg(Pbf)-OH and 20mmol HOBt in a 100mL beaker, cool to 5℃, add 35mL DMF solution and 3.1mL DIC, let it stand for 15min, and add the solution in the 100mL beaker to the above 250mL solid phase synthesis reactor. Stir and react for 1.5h. The reaction is complete; wash the resin with DMF solution 3 times, 70mL each time; after washing, proceed to the next step; add 70mL of 20% Pip / DMF (v / v) solution, stir and react for 30min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 70mL each time, and drain for use.

[0104] Take 20mmol Fmoc-D-Phe-OH and 20mmol HOBt in a 100mL beaker, cool to 5℃, add 35mL DMF solution and 3.1mL DIC, let it stand for 15min, and add the solution in the 100mL beaker to the above 250mL solid-phase synthesis reactor. Stir and react for 1.5h. The reaction is complete; wash the resin with DMF solution 3 times, 70mL each time; after washing, proceed to the next step; add 70mL of 20% Pip / DMF (v / v) solution, stir and react for 30min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 70mL each time, and drain for use.

[0105] 20 mmol Fmoc-His(Trt)-OH and 20 mmol HOBt were placed in a 100 mL beaker and cooled to 5°C. 35 mL of DMF solution and 20 mmol of DIC were added and allowed to react for 15 minutes. The solution in the 100 mL beaker was then added to the 250 mL solid-phase synthesis reactor described above and stirred for 1.5 hours. The reaction was completed. The resin was washed with DMF solution three times, 70 mL each time. After washing, the resin was processed to the next step. 70 mL of 20% Pip / DMF (v / v) solution was added and stirred for 30 minutes. The deprotection solution was removed by filtration and then washed with DMF solution six times, 40 mL each time, and dried. The resin was then washed with methanol twice, 70 mL each time, DCM twice, and methanol twice, 70 mL each time. The resin was dried under vacuum to obtain the peptide resin H-His(Trt)-D-Phe-Arg(Pbf)-Linker-AM resin. The peptide resin was cleaved with TFA / Tis / H2O (volume ratio of TFA, Tis and H2O was 90:5:5) in an amount of 100 mL for 2.5 h. The cleavage solution was added to 1000 mL of ether (5°C) solution to precipitate a white solid, which was centrifuged and dried in vacuo to obtain H-His-D-Phe-Arg-NH2. Its mass spectrum and LC-MS characterization results are shown in Figure 2. Figure 41 and Figure 42 As shown. A synthetic route of a hyaluronic acid modified beauty peptide is: ; Wherein, n=2, -COOX is -COONa; The preparation method of hyaluronic acid-modified cosmetic peptide comprises the following steps: Weigh 0.5g H-His-D-Phe-Arg-NH2.3TFA, add 5mL DMSO and stir to dissolve, add 661mg DIEA, then add 1.94g hyaluronic acid, control the temperature in a water bath at 45°C, stir and react overnight, take a sample and detect by LC-MS, the reaction is basically complete, add 4mL acetic acid, control the temperature in a water bath at 35°C, and carry out the rearrangement reaction for 3h, take a sample and detect by LC-MS, the reaction is basically complete; then perform reverse chromatography purification, purification conditions: Dissolution: dilute 0.5 g of crude product with 100 mL of H2O; Packing: 21.2*250mm, 10-120, C18; Flow rate: 10mL / min; Wavelength: 220nm; Mobile phase: A: 1% HAc; B: ACN; Balance: A:B=100:0, balance 10 min, flow rate: 10 mL / min; Sample loading: flow rate: 10 mL / min; Elution: 0-20%B 60 min; Column cleaning: 80% ACN cleaning to baseline equilibrium; Qualified products were collected to obtain three types of hyaluronic acid-modified beauty peptides, namely: Hyaluronic acid modified beauty peptide L2, in which n=0 in the structure, i.e. y=1 in the raw material hyaluronic acid structure, and the product prepared with the beauty peptide, its mass spectrometry and HPLC characterization results are as follows Figure 43 and Figure 44 As shown; Hyaluronic acid modified beauty peptide L4, in which n=1 in its structure, i.e. y=2 in the raw material hyaluronic acid structure, and the product prepared with the beauty peptide, its mass spectrometry and HPLC characterization results are as follows Figure 45 and Figure 46 As shown; Hyaluronic acid modified beauty peptide L6, in which n=2 in its structure, i.e. y=3 in the raw material hyaluronic acid structure, and the product prepared with the beauty peptide, its mass spectrometry and HPLC characterization results are as follows Figure 47 and Figure 48 shown.

[0106] Example 10: A method for synthesizing H-Tyr-Pro-Phe-Phe-NH2 comprises: Place 10 mmol of AM resin in a 250 mL solid-phase synthesis reactor, add 70 mL of 20% Pip / DMF (v / v) solution, stir and react for 30 min, filter to remove the deprotection solution, then wash with DMF solution 6 times, 70 mL each time, and drain to dryness for use.

[0107] Take 20mmol Fmoc-Linker and 20mmol HOBt in a 100mL beaker, cool to 5℃, add 35mL DMF solution and 3.1mL DIC, let it stand for 15min, and add the solution in the 100mL beaker to the above 250mL solid-phase synthesis reactor. Stir and react for 1.5h. The reaction is complete; wash the resin with DMF solution 3 times, 70mL each time; after washing, proceed to the next step; add 70mL of 20% Pip / DMF (v / v) solution, stir and react for 30min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 70mL each time, and drain and set aside.

[0108] Take 20mmol Fmoc-Phe-OH and 20mmol HOBt in a 100mL beaker, cool to 5℃, add 35mL of DMF solution and 3.1mL of DIC, let it stand for 15min, and add the solution in the 100mL beaker to the above 250mL solid-phase synthesis reactor. Stir and react for 1.5h. The reaction is complete; wash the resin with DMF solution 3 times, 70mL each time; after washing, proceed to the next step; add 70mL of 20% Pip / DMF (v / v) solution, stir and react for 30min, filter to remove the deprotection solution, and then wash with DMF solution L 6 times, 70mL each time, and drain for use.

[0109] Place 20 mmol of Fmoc-Phe-OH and 20 mmol of HOBt in a 100 mL beaker, cool to 5°C, add 35 mL of DMF solution and 3.1 mL of DIC, and allow to react for 15 minutes. The solution in the 100 mL beaker was then added to the 250 mL solid-phase synthesis reactor described above and stirred for 1.5 hours until the reaction was complete. Wash the resin three times with 70 mL of DMF solution each time. After washing, proceed to the next step. Add 70 mL of a 20% Pip / DMF (v / v) solution and stir for 30 minutes. Filter to remove the deprotection solution, then wash the resin six times with 70 mL of DMF solution each time and drain to dryness for later use.

[0110] Place 20 mmol of Fmoc-Pro-OH and 20 mmol of HOBt in a 100 mL beaker, cool to 5°C, add 35 mL of DMF solution and 3.1 mL of DIC, and allow to react for 15 minutes. The solution in the 100 mL beaker was then added to the 250 mL solid-phase synthesis reactor described above and stirred for 1.5 hours until the reaction was complete. Wash the resin three times with 70 mL of DMF solution each time. After washing, proceed to the next step. Add 70 mL of a 20% Pip / DMF (v / v) solution and stir for 30 minutes. Filter to remove the deprotection solution, then wash the resin six times with 70 mL of DMF solution each time and drain to dryness for later use.

[0111] Place 20 mmol of Fmoc-Tyr(tBu)-OH and 20 mmol of HOBt in a 100 mL beaker, cool to 5°C, add 35 mL of DMF solution and 3.1 mL of DIC, and allow to react for 15 minutes. The solution in the 100 mL beaker was then added to the 250 mL solid-phase synthesis reactor described above and stirred for 1.5 hours until the reaction was complete. The resin was washed three times with 70 mL of DMF solution. After washing, proceed to the next step. Add 70 mL of 20% Pip / DMF (v / v) solution and stir for 30 minutes. Filter to remove the deprotection solution, then wash six times with 40 mL of DMF solution and drain to dryness for later use. Then, wash with methanol twice (70 mL each), DCM twice (70 mL each), and methanol twice (70 mL each). Dry under vacuum to obtain the peptide resin H-Tyr(tBu)-Pro-Phe-Phe-Linker-AM resin. The peptide resin was cleaved with TFA / Tis / H2O (TFA, Tis, and H2O in a volume ratio of 90:5:5) in an amount of 100 mL for 2.5 h. The cleavage solution was added to 1000 mL of ether (5°C) to precipitate a white solid, which was centrifuged and dried in vacuo to obtain H-Tyr-Pro-Phe-Phe-NH2. Its mass spectrometry and HPLC characterization results are shown in FIG. Figure 49 and Figure 50 shown.

[0112] The preparation method of hyaluronic acid-modified cosmetic peptide comprises the following steps: Weigh 0.5g H-Tyr-Pro-Phe-Phe-NH2.TFA, add 5mL DMSO and stir to dissolve, add 0.771g DIEA, then add 2.27g hyaluronic acid, control the temperature in a water bath at 45°C, stir and react overnight, take a sample and detect by LC-MS, the reaction is basically complete, add 4mL acetic acid, control the temperature in a water bath at 35°C, and carry out the rearrangement reaction for 3h, take a sample and detect by LC-MS, the reaction is basically complete; then perform reverse phase chromatography purification, purification conditions: Dissolution: dilute 0.2 g of crude product with 100 mL of H2O; Packing: 21.2*250mm, 10-120, C18; Flow rate: 10mL / min; Wavelength: 220nm; Mobile phase: A: 1% HAc; B: ACN; Balance: A:B=95:5, balance 10 min, flow rate: 10 mL / min; Sample loading: flow rate: 10 mL / min; Elution: 12-32%B 60 min; Column cleaning: 80% ACN cleaning to baseline equilibrium; Qualified products were collected to obtain hyaluronic acid modified beauty peptides, and their mass spectrometry and HPLC characterization results were as follows: Figure 51 and Figure 52 shown.

[0113] Example 11: The preparation method of hyaluronic acid-modified cosmetic peptide comprises the following steps: Weigh 0.5g H-β-Ala-His-OH.TFA, add 10mL DMSO and stir to dissolve, add 0.5g TFA, stir for 10min, then add 2.5g DIEA, then add 6.85g hyaluronic acid, control the temperature in a water bath at 45°C, stir and react overnight, take a sample and detect LC-MS, the reaction is basically complete, add 4mL acetic acid, control the temperature in a water bath at 35°C, and carry out the rearrangement reaction for 3h, take a sample and detect LC-MS, the reaction is basically complete; then perform reverse chromatography purification, purification conditions: Dissolution: dilute 0.5 g of crude product with 100 mL of H2O; Packing: 21.2*250mm, 10-120, C18; Flow rate: 10mL / min; Wavelength: 220nm; Mobile phase: A: 1% HAc; B: ACN; Balance: A:B=100:0, balance 10 min, flow rate: 10 mL / min; Sample loading: flow rate: 10 mL / min; Elution: 0-10% B 60 min; Column cleaning: 80% ACN cleaning to baseline equilibrium; Qualified products were collected to prepare hyaluronic acid-modified cosmetic peptides, wherein the cosmetic peptides were combined with two tetrasaccharide sodium hyaluronates or the cosmetic peptides were combined with one hexasaccharide sodium hyaluronate and one disaccharide sodium hyaluronate. The mass spectrometry and HPLC characterization results of the products were as follows: Figure 53 and Figure 54As shown, the peak position of HPLC is 6.460min, and the molecular weight of the product is 872.3 / 582.0; the mass spectrum and HPLC characterization results of the product of the beauty peptide and a hexasaccharide sodium hyaluronate and a tetrasaccharide sodium hyaluronate are shown as follows Figure 55 and Figure 56 As shown, the peak position in the high performance liquid chromatography is 7.202 min, and the molecular weight of the product is 708.4 / 1061.8 / 1415.8.

[0114] Example 12: A method for synthesizing H-Glu-Glu-Met-Gln-Arg-Arg-Ala-OH, comprising: 7.5 mmol of Wang resin was placed in a 125 mL solid-phase synthesis reactor. 15 mmol of the amino acid Fmoc-Ala-OH was added, followed by 50 mL of dichloromethane, 4.8 mL of pyridine, and 4.2 mL of DBU. The mixture was allowed to react at 25°C for 3 h. The mixture was then filtered and washed with DMF (50 mL each) three times. A capping solution of Ac2O / DMF / DIEA (the volume ratio of Ac2O, DMF, and DIEA was 10:84:6) was added and allowed to react for 15 min. The mixture was then filtered and washed twice with dichloromethane (50 mL each), methanol (50 mL each), and DMF (50 mL each). A 20% Pip / DMF (v / v) solution (50 mL) was added and stirred for 30 min. The mixture was filtered to remove the deprotection solution and then washed six times with DMF (50 mL each). The mixture was then dried and used.

[0115] Place 15 mmol of Fmoc-Arg(Pbf)-OH and 15 mmol of HOBt in a 50 mL beaker, cool to 5°C, add 20 mL of DMF solution and 2.32 mL of DIC, and let the mixture react for 15 minutes. The mixture was then added to the 125 mL solid-phase synthesis reactor described above and stirred for 1.5 hours. The reaction was complete. The resin was washed three times with 15 mL of DMF solution. After washing, the resin was ready for the next step. 50 mL of a 20% Pip / DMF (v / v) solution was added and stirred for 30 minutes. The deprotection solution was removed by filtration, and the resin was then washed six times with 50 mL of DMF solution, drained, and set aside.

[0116] Place 15 mmol of Fmoc-Arg(Pbf)-OH and 15 mmol of HOBt in a 50 mL beaker, cool to 5°C, add 20 mL of DMF solution and 2.32 mL of DIC, and let the mixture react for 15 minutes. The mixture was then added to the 125 mL solid-phase synthesis reactor described above and stirred for 1.5 hours. The reaction was complete. The resin was washed three times with 15 mL of DMF solution. After washing, the resin was ready for the next step. 50 mL of a 20% Pip / DMF (v / v) solution was added and stirred for 30 minutes. The deprotection solution was removed by filtration, and the resin was then washed six times with 50 mL of DMF solution, drained, and set aside.

[0117] Place 15 mmol of Fmoc-Gln(Trt)-OH and 15 mmol of HOBt in a 50 mL beaker, cool to 5°C, add 20 mL of DMF solution and 2.32 mL of DIC, and allow to react for 15 minutes. The solution in the 50 mL beaker was then added to the 125 mL solid-phase synthesis reactor described above and stirred for 1.5 hours until the reaction was complete. The resin was washed three times with 15 mL of DMF solution. After washing, proceed to the next step. Add 50 mL of a 20% Pip / DMF (v / v) solution, stir, and react for 30 minutes. Filter to remove the deprotection solution, then wash the resin six times with 50 mL of DMF solution, drain, and set aside.

[0118] Place 15 mmol of Fmoc-Met-OH and 15 mmol of HOBt in a 50 mL beaker, cool to 5°C, add 20 mL of DMF solution and 2.32 mL of DIC, and let the mixture react for 15 minutes. The mixture was then added to the 125 mL solid-phase synthesis reactor described above and stirred for 1.5 hours. The reaction was complete. The resin was washed three times with 15 mL of DMF solution. After washing, the resin was ready for the next step. 50 mL of a 20% Pip / DMF (v / v) solution was added and stirred for 30 minutes. The deprotection solution was removed by filtration, and the resin was then washed six times with 50 mL of DMF solution. The mixture was then drained and used.

[0119] 15 mmol of Fmoc-Glu(OtBu)-OH and 15 mmol of HOBt were placed in a 50 mL beaker and cooled to 5°C. 20 mL of DMF solution and 2.32 mL of DIC were added and allowed to react for 15 minutes. The solution in the 50 mL beaker was then added to the 125 mL solid-phase synthesis reactor described above. The reaction was stirred for 1.5 hours until completion. The resin was washed three times with 15 mL of DMF solution. After washing, the resin was moved to the next step: 50 mL of a 20% Pip / DMF (v / v) solution was added and stirred for 30 minutes. The deprotection solution was removed by filtration and then washed six times with 50 mL of DMF solution. The resin was then dried and used.

[0120] 15 mmol of Fmoc-Glu(OtBu)-OH and 15 mmol of HOBt were placed in a 50 mL beaker and cooled to 5°C. 20 mL of DMF solution and 2.32 mL of DIC were added and allowed to react for 15 minutes. The solution in the 50 mL beaker was then added to the 125 mL solid-phase synthesis reactor described above and stirred for 1.5 hours until the reaction was complete. The resin was washed three times with 15 mL of DMF solution. After washing, the resin was moved to the next step: 50 mL of a 20% Pip / DMF (v / v) solution was added and stirred for 30 minutes. The deprotection solution was removed by filtration and then washed six times with 50 mL of DMF solution, two times with 50 mL of methanol, two times with 50 mL of DCM solution, and two times with 50 mL of methanol. Vacuum drying gave H-AA1-AA2-AA3-AA4-AA5-AA6-AA7-Wang-resin, wherein AA1 is Glu(OtBu); AA2 is Glu(OtBu); AA3 is Met; AA4 is Gln(Trt); AA5 is Arg(Pbf); AA6 is Arg(Pbf); and AA7 is Ala.

[0121] 4.22 g of the above peptide resin was cleaved with 40 mL of TFA / thioanisole / phenol / H2O / EDT (the mass ratio of TFA, thioanisole, phenol, H2O, and EDT was 87.5:5:2.5:2.5:2.5) for 2.5 h. The cleavage solution was added to 400 mL of ether (5°C) to precipitate a white solid, which was centrifuged and dried in vacuo to obtain H-Glu-Glu-Met-Gln-Arg-Arg-Ala-OH. Its mass spectrometry and HPLC characterization results are shown in Figure 2. Figure 57 and Figure 58 shown.

[0122] The preparation method of hyaluronic acid-modified cosmetic peptide comprises the following steps: Weigh 0.5 g H-Glu-Glu-Met-Gln-Arg-Arg-Ala-OH.3TFA, add 5 mL DMSO and stir to dissolve, then add 0.46 g DIEA and 1.24 g hyaluronic acid, control the temperature in a water bath at 45°C, stir and react overnight, take a sample and detect by LC-MS, if the reaction is basically complete, add 4 mL acetic acid, control the temperature in a water bath at 35°C, and carry out the rearrangement reaction for 3 hours, take a sample and detect by LC-MS, if the reaction is basically complete; then perform reverse chromatography purification, purification conditions: Dissolution: dilute 0.5 g of crude product with 100 mL of H2O; Packing: 21.2*250mm, 10-120, C18; Flow rate: 10mL / min; Wavelength: 220nm; Mobile phase: A: 1% HAc; B: ACN; Balance: A:B=100:0, balance 10 min, flow rate: 10 mL / min; Sample loading: flow rate: 10 mL / min; Elution: 0-10% B 60 min; Column cleaning: 80% ACN cleaning to baseline equilibrium; Qualified products were collected to obtain hyaluronic acid modified beauty peptides, and the HPLC test results were as follows: Figure 59 Among them, the sample at 11.577min is the product of the combination of the beauty peptide and a tetrasaccharide sodium hyaluronate, and its mass spectrometry characterization results are shown as follows. Figure 60 As shown; the sample at 12.223min is the product of the combination of the beauty peptide and a hexasaccharide sodium hyaluronate, and its mass spectrometry characterization results are shown as follows Figure 61 shown.

[0123] Example 13: A method for synthesizing H-Asp-Val-Lys-Tyr-OH, comprising: 6.25 mmol of CTC resin was placed in a 250 mL solid-phase synthesis reactor. 12.5 mmol of the amino acid Fmoc-Tyr(tBu)-OH was added, along with 120 mL of dichloromethane. The mixture was reacted at 25°C for 3 h, followed by 8.7 mL of DIEA. 12.5 mL of methanol was then added and allowed to react for 5 min. The mixture was filtered, and the resin was washed twice with 150 mL of dichloromethane, twice with 150 mL of methanol, and twice with 390 mL of DMF. 65 mL of a 20% Pip / DMF (v / v) solution was added, and the mixture was stirred for 30 min. The deprotection solution was removed by filtration, and the mixture was then washed six times with 120 mL of DMF solution before drying.

[0124] Place 15 mmol of Fmoc-Lys(Boc)-OH and 15 mmol of HOBt in a 100 mL beaker, cool to 5°C, add 14 mL of DMF solution and 1.89 mL of DIC, and allow to react for 15 minutes. The solution in the 100 mL beaker was then added to the 250 mL solid-phase synthesis reactor described above and stirred for 1.5 hours until the reaction was complete. The resin was washed three times with 120 mL of DMF solution. After washing, proceed to the next step. Add 65 mL of a 20% Pip / DMF (v / v) solution, stir, and react for 30 minutes. Filter to remove the deprotection solution, then wash the resin six times with 120 mL of DMF solution, drain, and set aside.

[0125] Place 15 mmol of Fmoc-Val-OH and 15 mmol of HOBt in a 100 mL beaker, cool to 5°C, add 10 mL of DMF solution and 2.32 mL of DIC, and let the mixture react for 15 minutes. The mixture was then added to the 250 mL solid-phase synthesis reactor described above and stirred for 1.5 hours. The reaction was complete. The resin was washed three times with 120 mL of DMF solution. After washing, the resin was ready for the next step. 65 mL of a 20% Pip / DMF (v / v) solution was added and stirred for 30 minutes. The deprotection solution was removed by filtration, and the resin was then washed six times with 120 mL of DMF solution. The mixture was then drained and used.

[0126] 15 mmol of Fmoc-Asp(OtBu)-OH and 15 mmol of HOBt were placed in a 100 mL beaker and cooled to 5°C. 13 mL of DMF solution and 2.32 mL of DIC were added and allowed to react for 15 minutes. The solution in the 100 mL beaker was then added to the 250 mL solid-phase synthesis reactor described above and stirred for 1.5 hours until the reaction was complete. The resin was washed three times with 120 mL of DMF solution. After washing, the resin was processed to the next step. 65 mL of 20% Pip / DMF (v / v) solution was added and stirred for 30 minutes. The deprotection solution was removed by filtration and then washed six times with 120 mL of DMF solution. The resin was then washed twice with 125 mL of methanol, twice with 125 mL of DCM solution, and twice with 125 mL of methanol. The resin was then dried under vacuum to obtain H-Asp(OtBu)-Val-Lys(Boc)-Tyr(tBu)-CTC-resin. 4.89 g of the peptide resin was cleaved with TFA / Tis / H2O (TFA, Tis, and H2O in a volume ratio of 90:5:5) using 40 mL of solution. The mixture was stirred at 30°C for 2.5 h, filtered, and the resin removed to obtain a filtrate. The filtrate was dried to obtain the crude peptide H-Asp-Val-Lys-Tyr-OH. The mass spectrometry and HPLC characterization results are shown in the following table. Figure 62 and Figure 63 shown.

[0127] The preparation method of hyaluronic acid-modified cosmetic peptide comprises the following steps: Weigh 0.5g H-Asp-Val-Lys-Tyr-OH.2TFA, add 7mL DMSO and stir to dissolve, then add 0.773g DIEA, then add 2.1g hyaluronic acid, control the temperature in a water bath at 45°C, stir for 15 minutes to dissolve, then stir and react overnight, take a sample and detect by LC-MS, if the reaction is basically complete, add 4mL acetic acid, control the temperature in a water bath at 35°C, and carry out the rearrangement reaction for 3 hours, take a sample and detect by LC-MS, if the reaction is basically complete; then perform reverse chromatography purification, purification conditions: Dissolution: dilute 0.5 g of crude product with 100 mL of H2O; Packing: 21.2*250mm, 10-120, C18; Flow rate: 10mL / min; Wavelength: 220nm; Mobile phase: A: 1% HAc; B: ACN; Balance: A:B=100:0, balance 10 min, flow rate: 10 mL / min; Sample loading: flow rate: 10 mL / min; Elution: 0-20%B 60 min; Column cleaning: 80% ACN cleaning to baseline equilibrium; Qualified products were collected to obtain hyaluronic acid modified beauty peptides, and the HPLC test results were as follows: Figure 64 Among them, the sample at 8.226min is the product of the combination of the beauty peptide and two hexasaccharide sodium hyaluronates, and its mass spectrometry characterization results are shown as follows. Figure 65 As shown; the sample at 12.711min is the product of the combination of the beauty peptide and a hexasaccharide sodium hyaluronate, a disaccharide sodium hyaluronate, or the combination of the beauty peptide and two tetrasaccharide sodium hyaluronates. The mass spectrometry characterization results are shown as follows Figure 66 shown.

[0128] Example 14: A method for synthesizing H-His-Ala-Leu-Arg-Phe-Trp-NH2, comprising: Place 30 mmol of AM resin in a 500 mL solid-phase synthesis reactor, add 200 mL of 20% Pip / DMF (v / v) solution, stir and react for 30 min, filter to remove the deprotection solution, then wash with DMF solution 6 times, 200 mL each time, and drain to set aside.

[0129] Place 10 mmol of Fmoc-Linker and 60 mmol of HOBt in a 100 mL beaker, cool to 5°C, add 60 mL of DMF solution and 9.3 mL of DIC, and allow to react for 15 minutes. The solution in the 100 mL beaker was then added to the 500 mL solid-phase synthesis reactor described above and stirred for 1.5 hours until the reaction was complete. The resin was washed three times with 100 mL of DMF solution. After washing, proceed to the next step: add 200 mL of 20% Pip / DMF (v / v) solution, stir for 30 minutes, filter to remove the deprotection solution, and then wash six times with 200 mL of DMF solution. Drain and set aside.

[0130] Place 60 mmol of Fmoc-Trp(Boc)-OH and 60 mmol of HOBt in a 100 mL beaker, cool to 5°C, add 60 mL of DMF solution and 9.3 mL of DIC, and allow to react for 15 minutes. The solution in the 100 mL beaker was then added to the 500 mL solid-phase synthesis reactor described above and stirred for 1.5 hours until the reaction was complete. The resin was washed three times with 100 mL of DMF solution. After washing, proceed to the next step. Add 200 mL of a 20% Pip / DMF (v / v) solution and stir for 30 minutes. Filter to remove the deprotection solution, then wash the resin six times with 200 mL of DMF solution, draining to dryness.

[0131] Place 60 mmol of Fmoc-Phe-OH and 60 mmol of HOBt in a 100 mL beaker, cool to 5°C, add 60 mL of DMF solution and 9.3 mL of DIC, and allow to react for 15 minutes. The solution in the 100 mL beaker was then added to the 500 mL solid-phase synthesis reactor described above and stirred for 1.5 hours until the reaction was complete. The resin was washed three times with 100 mL of DMF solution. After washing, proceed to the next step. Add 200 mL of a 20% Pip / DMF (v / v) solution and stir for 30 minutes. Filter to remove the deprotection solution, then wash the resin six times with 200 mL of DMF solution, draining thoroughly and setting aside.

[0132] Place 60 mmol of Fmoc-Arg(Pbf)-OH and 60 mmol of HOBt in a 100 mL beaker, cool to 5°C, add 60 mL of DMF solution and 9.3 mL of DIC, and allow to react for 15 minutes. The solution in the 100 mL beaker was then added to the 500 mL solid-phase synthesis reactor described above and stirred for 1.5 hours until the reaction was complete. The resin was washed three times with 100 mL of DMF solution. After washing, proceed to the next step. Add 200 mL of a 20% Pip / DMF (v / v) solution and stir for 30 minutes. Filter to remove the deprotection solution, then wash the resin six times with 200 mL of DMF solution, draining to dryness.

[0133] Place 60 mmol of Fmoc-Leu-OH and 60 mmol of HOBt in a 100 mL beaker, cool to 5°C, add 60 mL of DMF solution and 9.3 mL of DIC, and allow to react for 15 minutes. The solution in the 100 mL beaker was then added to the 500 mL solid-phase synthesis reactor described above and stirred for 1.5 hours until the reaction was complete. Wash the resin three times with 100 mL of DMF solution. After washing, proceed to the next step. Add 200 mL of a 20% Pip / DMF (v / v) solution and stir for 30 minutes. Filter to remove the deprotection solution, then wash the resin six times with 200 mL of DMF solution, draining to dryness.

[0134] Place 60 mmol of Fmoc-Ala-OH and 60 mmol of HOBt in a 100 mL beaker, cool to 5°C, add 60 mL of DMF solution and 9.3 mL of DIC, and allow to react for 15 minutes. The solution in the 100 mL beaker was then added to the 500 mL solid-phase synthesis reactor described above and stirred for 1.5 hours until the reaction was complete. Wash the resin three times with 100 mL of DMF solution each time. After washing, proceed to the next step. Add 200 mL of a 20% Pip / DMF (v / v) solution and stir for 30 minutes. Filter to remove the deprotection solution, then wash the resin six times with 200 mL of DMF solution each time, drain, and set aside.

[0135] 90 mmol of Fmoc-His(Trt)-OH and 90 mmol of HOBt were placed in a 100 mL beaker and cooled to 5°C. 100 mL of DMF solution and 13.9 mL of DIC were added and allowed to react for 15 minutes. The solution in the 100 mL beaker was then added to a 500 mL solid-phase synthesis reactor and stirred for 1.5 hours until the reaction was complete. The resin was washed three times with 150 mL of DMF solution. After washing, the resin was processed for the next step. 200 mL of a 20% Pip / DMF (v / v) solution was added and stirred for 30 minutes. The deprotection solution was removed by filtration and then washed six times with 200 mL of DMF solution. The mixture was then dried and set aside. The resin was then washed twice with 200 mL of methanol, twice with 200 mL of DCM solution, and twice with 200 mL of methanol. Drying under vacuum gave a peptide resin of H-AA1-AA2-AA3-AA4-AA5-AA6-Linker-AM resin, wherein AA1 is His (Trt); AA2 is Ala; AA3 is Leu; AA4 is Arg (Pbf); AA5 is Phe; and AA6 is Trp.

[0136] The peptide resin was cleaved with 690 mL of TFA / thioanisole / phenol / H2O / EDT (the mass ratio of TFA, thioanisole, phenol, H2O and EDT was 87.5:5:2.5:2.5:2.5) for 2.5 h. The cleavage solution was added to 7000 mL of ether (5°C) to precipitate a white solid, which was centrifuged and dried in vacuo to obtain H-His-Ala-Leu-Arg-Phe-Trp-NH2. Its mass spectrometry and HPLC characterization results are shown in Figure 2. Figure 67 and Figure 68 shown.

[0137] A synthetic route for a hyaluronic acid-modified beauty peptide is: ; Wherein, -COOX is -COONa; The preparation method of hyaluronic acid-modified cosmetic peptide comprises the following steps: Weigh 1g H-His-Ala-Leu-Arg-Phe-Trp-NH2.TFA, add 10mL DMSO and stir to dissolve, add 1.28g DIEA, stir for 5 minutes, then add 3.75g hyaluronic acid, control the temperature in a water bath at 45°C, stir and react overnight, take a sample and detect LC-MS, the reaction is basically complete, add 8mL acetic acid, control the temperature in a water bath at 35°C, and carry out the rearrangement reaction for 3h, take a sample and detect LC-MS, the reaction is basically complete; then perform reverse chromatography purification, purification conditions: Dissolution: dilute 0.2 g of crude product with 100 mL of H2O; Packing: 21.2*250mm, 10-120, C18; Flow rate: 10mL / min; Wavelength: 220nm; Mobile phase: A: 1% HAc; B: ACN; Balance: A:B=95:5, balance 10 min, flow rate: 10 mL / min; Sample loading: flow rate: 10 mL / min; Elution: 12-32%B 60 min; Column cleaning: 80% ACN cleaning to baseline equilibrium; Qualified products were collected to obtain three structures of hyaluronic acid-modified beauty peptides, as shown below: A2, its mass spectrometry and HPLC characterization results are as follows Figure 69 and Figure 70 As shown; A4, its mass spectrometry and HPLC characterization results are as follows Figure 71 and Figure 72 Its H NMR spectrum is shown as Figures 73-1 to 73-5As shown, and the carbon spectrum is as Figures 74-1 to 74-5 As shown, its two-dimensional cosy map is as follows Figure 75 As shown; A6, its mass spectrometry and HPLC characterization results are as follows Figure 76 and Figure 77 shown.

[0138] Example 15: The preparation method of hyaluronic acid-modified cosmetic peptide comprises the following steps: Weigh 0.5g H-β-Ala-Pro-Dab-NH-Bzl.TFA, add 5mL DMSO and stir to dissolve, add 704.32mg DIEA, stir for 5min, then add 2064.08mg hyaluronic acid, control the temperature in a water bath at 45°C, stir and react overnight, take a sample and detect by LC-MS, the reaction is basically complete, add 3mL acetic acid, control the temperature in a water bath at 35°C, and carry out the rearrangement reaction for 3h, take a sample and detect by LC-MS, the reaction is basically complete; then perform reverse phase chromatography purification, purification conditions: Dissolution: dilute 0.2 g of crude product with 100 mL of H2O; Packing: 21.2*250mm, 10-120, C18; Flow rate: 10mL / min; Wavelength: 220nm; Mobile phase: A: 1% HAc; B: ACN; Balance: A:B=95:5, balance 10 min, flow rate: 10 mL / min; Sample loading: flow rate: 10 mL / min; Elution: 12-32%B 60 min; Column cleaning: 80% ACN cleaning to baseline equilibrium; Qualified products are collected to prepare hyaluronic acid-modified beauty peptides.

[0139] Example 16: The preparation method of hyaluronic acid-modified cosmetic peptide comprises the following steps: Weigh 0.5 g of H-Dab-Val-Dab-OH.2TFA, add 7 mL of DMSO and stir to dissolve, then add 833.2 mg of DIEA and stir for 5 minutes, then add 2441.79 mg of hyaluronic acid, control the temperature in a water bath at 45°C, and stir to react overnight. Samples were taken for LC-MS analysis, and the reaction was essentially complete. 3 mL of acetic acid was added, and the temperature in a water bath was controlled at 35°C. The rearrangement reaction was carried out for 3 hours, and samples were taken for LC-MS analysis, and the reaction was essentially complete. Then, reverse chromatography purification was performed under the following purification conditions: Dissolution: dilute 0.5 g of crude product with 100 mL of H2O; Packing: 21.2*250mm, 10-120, C18; Flow rate: 10mL / min; Wavelength: 220nm; Mobile phase: A: 1% HAc; B: ACN; Balance: A:B=100:0, balance 10 min, flow rate: 10 mL / min; Sample loading: flow rate: 10 mL / min; Elution: 0-20%B 60 min; Column cleaning: 80% ACN cleaning to baseline equilibrium; Qualified products are collected to prepare hyaluronic acid-modified beauty peptides.

[0140] Example 17: The preparation method of hyaluronic acid-modified cosmetic peptide comprises the following steps: Weigh 0.5g H-Lys-Val-Lys-OH.2TFA, add 7mL DMSO and stir to dissolve, then add 708.03mg DIEA, stir for 5min, then add 2074.97mg hyaluronic acid, control the temperature in a water bath at 45°C, and stir to react overnight. Samples were taken for LC-MS analysis. If the reaction was essentially complete, 3mL acetic acid was added, and the temperature in a water bath was controlled at 35°C. The rearrangement reaction was carried out for 3h. Samples were taken for LC-MS analysis. If the reaction was essentially complete, the product was purified by reversed-phase chromatography. Purification conditions were as follows: Dissolution: dilute 0.5 g of crude product with 100 mL of H2O; Packing: 21.2*250mm, 10-120, C18; Flow rate: 10mL / min; Wavelength: 220nm; Mobile phase: A: 1% HAc; B: ACN; Balance: A:B=100:0, balance 10 min, flow rate: 10 mL / min; Sample loading: flow rate: 10 mL / min; Elution: 0-20%B 60 min; Column cleaning: 80% ACN cleaning to baseline equilibrium; Qualified products are collected to prepare hyaluronic acid-modified beauty peptides.

[0141] Example 18: The preparation method of hyaluronic acid-modified cosmetic peptide comprises the following steps: Weigh 0.5g H-Gln-Asp-Val-His-OH.TFA, add 5mL DMSO and stir to dissolve, add 531.55mg DIEA, stir for 5 minutes, then add 1557.76mg hyaluronic acid, control the temperature in a water bath at 45°C, stir and react overnight, take a sample and detect LC-MS, the reaction is basically complete, add 3mL acetic acid, control the temperature in a water bath at 35°C, and carry out the rearrangement reaction for 3h, take a sample and detect LC-MS, the reaction is basically complete; then perform reverse chromatography purification, purification conditions: Dissolution: dilute 0.2 g of crude product with 100 mL of H2O; Packing: 21.2*250mm, 10-120, C18; Flow rate: 10mL / min; Wavelength: 220nm; Mobile phase: A: 1% HAc; B: ACN; Balance: A:B=95:5, balance 10 min, flow rate: 10 mL / min; Sample loading: flow rate: 10 mL / min; Elution: 12-32%B 60 min; Column cleaning: 80% ACN cleaning to baseline equilibrium; Qualified products are collected to prepare hyaluronic acid-modified beauty peptides.

[0142] Example 19: The preparation method of hyaluronic acid-modified cosmetic peptide comprises the following steps: Weigh 0.5g H-Pro-Pro-Tyr-Leu-OH.TFA, add 5mL DMSO and stir to dissolve, add 541.25mg DIEA, stir for 5 minutes, then add 1586.20mg hyaluronic acid, control the temperature in a water bath at 45°C, stir and react overnight, take a sample and detect by LC-MS, the reaction is basically complete, add 3mL acetic acid, control the temperature in a water bath at 35°C, and carry out the rearrangement reaction for 3 hours, take a sample and detect by LC-MS, the reaction is basically complete; then perform reverse chromatography purification, purification conditions: Dissolution: dilute 0.2 g of crude product with 100 mL of H2O; Packing: 21.2*250mm, 10-120, C18; Flow rate: 10mL / min; Wavelength: 220nm; Mobile phase: A: 1% HAc; B: ACN; Balance: A:B=95:5, balance 10 min, flow rate: 10 mL / min; Sample loading: flow rate: 10 mL / min; Elution: 12-32%B 60 min; Column cleaning: 80% ACN cleaning to baseline equilibrium; Qualified products are collected to prepare hyaluronic acid-modified beauty peptides.

[0143] Example 20: The preparation method of hyaluronic acid-modified cosmetic peptide comprises the following steps: Weigh 0.5g H-Pro-Lys-Glu-Lys-OH.2TFA, add 7mL DMSO and stir to dissolve, then add 528.27mg DIEA, stir for 5min, then add 1548.14mg hyaluronic acid, control the temperature in a water bath at 45°C, and stir to react overnight. Samples were taken for LC-MS analysis. If the reaction was essentially complete, 3mL acetic acid was added, and the temperature in a water bath was controlled at 35°C. The rearrangement reaction was carried out for 3h. Samples were taken for LC-MS analysis. If the reaction was essentially complete, the product was purified by reversed-phase chromatography. Purification conditions were as follows: Dissolution: dilute 0.5 g of crude product with 100 mL of H2O; Packing: 21.2*250mm, 10-120, C18; Flow rate: 10mL / min; Wavelength: 220nm; Mobile phase: A: 1% HAc; B: ACN; Balance: A:B=100:0, balance 10 min, flow rate: 10 mL / min; Sample loading: flow rate: 10 mL / min; Elution: 0-20%B 60 min; Column cleaning: 80% ACN cleaning to baseline equilibrium; Qualified products are collected to prepare hyaluronic acid-modified beauty peptides.

[0144] Example 21: The preparation method of hyaluronic acid-modified cosmetic peptide comprises the following steps: Weigh 0.5g H-Val-Trp-OH.2TFA, add 7mL DMSO and stir to dissolve, then add 871.74mg DIEA, stir for 5 minutes, then add 2554.72mg hyaluronic acid, control the temperature in a water bath at 45°C, and stir to react overnight. Samples were taken for LC-MS analysis. If the reaction was essentially complete, 3mL acetic acid was added, and the temperature in a water bath was controlled at 35°C. The rearrangement reaction was carried out for 3 hours. Samples were taken for LC-MS analysis. If the reaction was essentially complete, the product was purified by reversed-phase chromatography. Purification conditions were as follows: Dissolution: dilute 0.5 g of crude product with 100 mL of H2O; Packing: 21.2*250mm, 10-120, C18; Flow rate: 10mL / min; Wavelength: 220nm; Mobile phase: A: 1% HAc; B: ACN; Balance: A:B=100:0, balance 10 min, flow rate: 10 mL / min; Sample loading: flow rate: 10 mL / min; Elution: 0-20%B 60 min; Column cleaning: 80% ACN cleaning to baseline equilibrium; Qualified products are collected to prepare hyaluronic acid-modified beauty peptides.

[0145] Example 22: The preparation method of hyaluronic acid-modified cosmetic peptide comprises the following steps: Weigh 0.5 g of H-Ser-Val-Val-Val-Arg-Thr-NH2.TFA, add 5 mL of DMSO and stir to dissolve, add 404.41 mg of DIEA, stir for 5 minutes, then add 1176.38 mg of hyaluronic acid, control the temperature in a water bath at 45°C, stir and react overnight, take a sample and detect by LC-MS, if the reaction is basically complete, add 3 mL of acetic acid, control the temperature in a water bath at 35°C, and carry out the rearrangement reaction for 3 hours, take a sample and detect by LC-MS, if the reaction is basically complete; then perform reverse chromatography purification, purification conditions: Dissolution: dilute 0.2 g of crude product with 100 mL of H2O; Packing: 21.2*250mm, 10-120, C18; Flow rate: 10mL / min; Wavelength: 220nm; Mobile phase: A: 1% HAc; B: ACN; Balance: A:B=95:5, balance 10 min, flow rate: 10 mL / min; Sample loading: flow rate: 10 mL / min; Elution: 12-32%B 60 min; Column cleaning: 80% ACN cleaning to baseline equilibrium; Qualified products are collected to prepare hyaluronic acid-modified beauty peptides.

[0146] Example 23: The preparation method of hyaluronic acid-modified cosmetic peptide comprises the following steps: Weigh 0.5g of Pal-Lys-Met(O)2-Lys-OH.2TFA, add 7mL of DMSO and stir to dissolve, then add 391.22mg of DIEA, stir for 5 minutes, then add 1146.50mg of hyaluronic acid, control the temperature in a water bath at 45°C, and stir to react overnight. Samples were taken for LC-MS analysis, and the reaction was basically complete. Add 3mL of acetic acid, control the temperature in a water bath at 35°C, and carry out the rearrangement reaction for 3 hours. Samples were taken for LC-MS analysis, and the reaction was basically complete. Then, reverse chromatography purification was performed. Purification conditions: Dissolution: dilute 0.5 g of crude product with 100 mL of H2O; Packing: 21.2*250mm, 10-120, C18; Flow rate: 10mL / min; Wavelength: 220nm; Mobile phase: A: 1% HAc; B: ACN; Balance: A:B=100:0, balance 10 min, flow rate: 10 mL / min; Sample loading: flow rate: 10 mL / min; Elution: 0-20%B 60 min; Column cleaning: 80% ACN cleaning to baseline equilibrium; Qualified products are collected to prepare hyaluronic acid-modified beauty peptides.

[0147] Example 24: The preparation method of hyaluronic acid-modified cosmetic peptide comprises the following steps: Weigh 0.5g H-Lys-Met(O)2-Lys-OH.2TFA, add 7mL DMSO and stir to dissolve, then add 604.37mg DIEA, stir for 5min, then add 1771.19mg hyaluronic acid, control the temperature in a water bath at 45°C, and stir to react overnight. Samples were taken for LC-MS detection. If the reaction was basically complete, 3mL acetic acid was added, and the temperature in a water bath was controlled at 35°C. The rearrangement reaction was carried out for 3h. Samples were taken for LC-MS detection. If the reaction was basically complete, the product was purified by reverse chromatography. The purification conditions were as follows: Dissolution: dilute 0.5 g of crude product with 100 mL of H2O; Packing: 21.2*250mm, 10-120, C18; Flow rate: 10mL / min; Wavelength: 220nm; Mobile phase: A: 1% HAc; B: ACN; Balance: A:B=100:0, balance 10 min, flow rate: 10 mL / min; Sample loading: flow rate: 10 mL / min; Elution: 0-20%B 60 min; Column cleaning: 80% ACN cleaning to baseline equilibrium; Qualified products are collected to prepare hyaluronic acid-modified beauty peptides.

[0148] Example 25: The preparation method of hyaluronic acid-modified cosmetic peptide comprises the following steps: Weigh 0.5 g of H-Val-Gly-Val-Ala-Pro-Gly-OH.2TFA, add 7 mL of DMSO and stir to dissolve, then add 530.41 mg of DIEA, stir for 5 minutes, then add 1554.41 mg of hyaluronic acid, control the temperature in a water bath at 45°C, and stir to react overnight. Samples were taken for LC-MS analysis, and the reaction was basically complete. 3 mL of acetic acid was added, and the temperature in a water bath was controlled at 35°C. The rearrangement reaction was carried out for 3 hours, and samples were taken for LC-MS analysis, and the reaction was basically complete. Then, reverse chromatography purification was performed. Purification conditions: Dissolution: dilute 0.5 g of crude product with 100 mL of H2O; Packing: 21.2*250mm, 10-120, C18; Flow rate: 10mL / min; Wavelength: 220nm; Mobile phase: A: 1% HAc; B: ACN; Balance: A:B=100:0, balance 10 min, flow rate: 10 mL / min; Sample loading: flow rate: 10 mL / min; Elution: 0-20%B 60 min; Column cleaning: 80% ACN cleaning to baseline equilibrium; Qualified products are collected to prepare hyaluronic acid-modified beauty peptides.

[0149] Example 26: The preparation method of hyaluronic acid-modified cosmetic peptide comprises the following steps: Weigh 0.5 g H-bataAla-His-Ser-His-OH.2TFA, add 7 mL DMSO and stir to dissolve, then add 587.07 mg DIEA, stir for 5 minutes, then add 1720.46 mg hyaluronic acid, control the temperature in a water bath at 45°C, and stir to react overnight. Samples were taken for LC-MS analysis, and the reaction was essentially complete. 3 mL acetic acid was added, and the temperature in a water bath was controlled at 35°C. The rearrangement reaction was carried out for 3 hours, and samples were taken for LC-MS analysis, and the reaction was essentially complete. Then, reverse chromatography purification was performed. Purification conditions: Dissolution: dilute 0.5 g of crude product with 100 mL of H2O; Packing: 21.2*250mm, 10-120, C18; Flow rate: 10mL / min; Wavelength: 220nm; Mobile phase: A: 1% HAc; B: ACN; Balance: A:B=100:0, balance 10 min, flow rate: 10 mL / min; Sample loading: flow rate: 10 mL / min; Elution: 0-20%B 60 min; Column cleaning: 80% ACN cleaning to baseline equilibrium; Qualified products are collected to prepare hyaluronic acid-modified beauty peptides.

[0150] Example 27: A method for synthesizing H-Trp-Phe-Arg-D-Leu-Ala-His-NH2 comprises: Place 5 mmol AM resin in a 100 mL solid-phase synthesis reactor, add 30 mL of 20% Pip / DMF (v / v) solution, stir and react for 30 min, filter to remove the deprotection solution, then wash with DMF solution 6 times, 30 mL each time, and drain to set aside.

[0151] Place 10 mmol of Fmoc-Linker and 10 mmol of HOBt in a 100 mL beaker, cool to 5°C, add 15 mL of DMF solution and 1.5 mL of DIC, and allow to react for 15 minutes. The solution in the 100 mL beaker was then added to the 100 mL solid-phase synthesis reactor described above and stirred for 1.5 hours until the reaction was complete. The resin was washed three times with 20 mL of DMF solution. After washing, proceed to the next step: add 30 mL of 20% Pip / DMF (v / v) solution, stir for 30 minutes, filter to remove the deprotection solution, and then wash six times with 30 mL of DMF solution before draining.

[0152] Place 10 mmol of Fmoc-His(Trt)-OH and 10 mmol of HOBt in a 100 mL beaker, cool to 5°C, add 15 mL of DMF solution and 1.5 mL of DIC, and allow to react for 15 minutes. The solution in the 100 mL beaker was then added to the 100 mL solid-phase synthesis reactor described above and stirred for 1.5 hours until the reaction was complete. Wash the resin three times with 20 mL of DMF solution each time. After washing, proceed to the next step. Add 30 mL of a 20% Pip / DMF (v / v) solution, stir, and react for 30 minutes. Filter to remove the deprotection solution, then wash the resin six times with 30 mL of DMF solution each time, drain, and set aside.

[0153] Place 10 mmol of Fmoc-Ala-OH and 10 mmol of HOBt in a 100 mL beaker, cool to 5°C, add 15 mL of DMF solution and 1.5 mL of DIC, and let the mixture react for 15 minutes. The solution in the 100 mL beaker was then added to the 100 mL solid-phase synthesis reactor described above. Stir and react for 1.5 hours until the reaction is complete. Wash the resin three times with 20 mL of DMF solution. After washing, proceed to the next step. Add 30 mL of 20% Pip / DMF (v / v) solution, stir and react for 30 minutes, filter to remove the deprotection solution, and then wash the resin six times with 30 mL of DMF solution. Drain and set aside.

[0154] Place 10 mmol of Fmoc-D-Leu-OH and 10 mmol of HOBt in a 100 mL beaker, cool to 5°C, add 15 mL of DMF solution and 1.5 mL of DIC, and allow to react for 15 minutes. The solution in the 100 mL beaker was then added to the 100 mL solid-phase synthesis reactor described above and stirred for 1.5 hours until the reaction was complete. Wash the resin three times with 20 mL of DMF solution each time. After washing, proceed to the next step. Add 30 mL of 20% Pip / DMF (v / v) solution, stir for 30 minutes, filter to remove the deprotection solution, and then wash the resin six times with 30 mL of DMF solution each time. Drain and set aside.

[0155] Place 10 mmol of Fmoc-Arg(Pbf)-OH and 10 mmol of HOBt in a 100 mL beaker, cool to 5°C, add 15 mL of DMF solution and 1.5 mL of DIC, and allow to react for 15 minutes. The solution in the 100 mL beaker was then added to the 100 mL solid-phase synthesis reactor described above and stirred for 1.5 hours until the reaction was complete. The resin was washed three times with 20 mL of DMF solution. After washing, proceed to the next step. Add 40 mL of a 20% Pip / DMF (v / v) solution, stir, and react for 30 minutes. Filter to remove the deprotection solution, then wash the resin six times with 40 mL of DMF solution, drain, and set aside.

[0156] Take 10mmol Fmoc-Phe-OH and 10mmol HOBt in a 100mL beaker, cool to 5℃, add 15mL of DMF solution and 1.5mL of DIC, let it stand for 15min, and add the solution in the 100mL beaker to the above 100mL solid phase synthesis reactor, stir and react for 1.5h, and the reaction is complete. Wash the resin with DMF solution 3 times, 20mL each time. After washing, proceed to the next step of the reaction. Add 40mL of 20% Pip / DMF (v / v) solution, stir and react for 30min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 40mL each time, and drain for use. 15 mmol of Fmoc-Trp(Boc)-OH and 1 mmol of HOBt were placed in a 100 mL beaker and cooled to 5°C. 15 mL of DMF solution and 2.3 mL of DIC were added and allowed to react for 15 minutes. The solution in the 100 mL beaker was then added to the 100 mL solid-phase synthesis reactor described above and stirred for 1.5 hours until the reaction was complete. The resin was washed three times with 30 mL of DMF solution. After washing, the resin was proceeded to the next step. 40 mL of a 20% Pip / DMF (v / v) solution was added and stirred for 30 minutes. The deprotection solution was removed by filtration and then washed six times with 40 mL of DMF solution. The mixture was then dried and set aside. The resin was then washed twice with 40 mL of methanol, twice with 40 mL of DCM solution, and twice with 40 mL of methanol. Drying under vacuum gave a peptide resin of H-AA1-AA2-AA3-AA4-AA5-AA6-Linker-AM resin, wherein AA1 is Trp(Boc); AA2 is Phe; AA3 is Arg; AA4 is D-Leu; AA5 is Ala; and AA6 is His(Trt).

[0157] The peptide resin was cleaved with 110 mL of TFA / thioanisole / phenol / H2O / EDT (the mass ratio of TFA, thioanisole, phenol, H2O and EDT was 87.5:5:2.5:2.5:2.5) for 2.5 h. The cleavage solution was added to 1000 mL of ether (5°C) to precipitate a white solid, which was centrifuged and dried in vacuo to obtain H-Trp-Phe-Arg-D-Leu-Ala-His-NH2. Its mass spectrometry and HPLC characterization results are shown in Figure 2. Figure 78 and Figure 79 shown.

[0158] A synthetic route for a hyaluronic acid-modified beauty peptide is: ; Wherein, -COOX is -COONa; The preparation method of hyaluronic acid-modified cosmetic peptide comprises the following steps: Weigh 1g of H-Trp-Phe-Arg-D-Leu-Ala-His-NH2.TFA, add 10mL of DMSO and stir to dissolve, add 1.28g of DIEA, stir for 5 minutes, then add 3.75g of hyaluronic acid, control the temperature in a water bath at 45°C, stir and react overnight, take a sample and detect by LC-MS, if the reaction is basically complete, add 8mL of acetic acid, keep the temperature constant for rearrangement reaction for 2h, take a sample and detect by LC-MS, if the reaction is basically complete; then perform reverse chromatography purification, purification conditions: Dissolution: dilute 0.2 g of crude product with 100 mL of H2O; Packing: 21.2*250mm, 10-120, C18; Flow rate: 10mL / min; Wavelength: 220nm; Mobile phase: A: 1% HAc; B: ACN; Balance: A:B=95:5, balance 10 min, flow rate: 10 mL / min; Sample loading: flow rate: 10 mL / min; Elution: 12-32%B 60 min; Column cleaning: 80% ACN cleaning to baseline equilibrium; Qualified products were collected to obtain three structures of hyaluronic acid-modified beauty peptides, as shown below: E2, its mass spectrometry and HPLC characterization results are shown in Figure 80 and Figure 81 As shown; E4, its mass spectrometry and HPLC characterization results are shown in Figure 82 and Figure 83 As shown; E6, its mass spectrometry and HPLC characterization results are shown in Figure 84 and Figure 85 shown.

[0159] Example 28: A method for synthesizing H-Trp-Phe-Arg-Leu-Ala-His-NH2 comprises: Place 5 mmol AM resin in a 100 mL solid-phase synthesis reactor, add 30 mL of 20% Pip / DMF (v / v) solution, stir and react for 30 min, filter to remove the deprotection solution, then wash with DMF solution 6 times, 30 mL each time, and drain to set aside.

[0160] Place 10 mmol of Fmoc-Linker and 10 mmol of HOBt in a 100 mL beaker, cool to 5°C, add 15 mL of DMF solution and 1.5 mL of DIC, and allow to react for 15 minutes. The solution in the 100 mL beaker was then added to the 100 mL solid-phase synthesis reactor described above and stirred for 1.5 hours until the reaction was complete. The resin was washed three times with 20 mL of DMF solution. After washing, proceed to the next step: add 30 mL of 20% Pip / DMF (v / v) solution, stir for 30 minutes, filter to remove the deprotection solution, and then wash six times with 30 mL of DMF solution before draining.

[0161] Place 10 mmol of Fmoc-His(Trt)-OH and 10 mmol of HOBt in a 100 mL beaker, cool to 5°C, add 15 mL of DMF solution and 1.5 mL of DIC, and allow to react for 15 minutes. The solution in the 100 mL beaker was then added to the 100 mL solid-phase synthesis reactor described above and stirred for 1.5 hours until the reaction was complete. Wash the resin three times with 20 mL of DMF solution each time. After washing, proceed to the next step. Add 30 mL of a 20% Pip / DMF (v / v) solution, stir, and react for 30 minutes. Filter to remove the deprotection solution, then wash the resin six times with 30 mL of DMF solution each time, drain, and set aside.

[0162] Place 10 mmol of Fmoc-Ala-OH and 10 mmol of HOBt in a 100 mL beaker, cool to 5°C, add 15 mL of DMF solution and 1.5 mL of DIC, and allow to react for 15 minutes. The solution in the 100 mL beaker was then added to the 100 mL solid-phase synthesis reactor described above and stirred for 1.5 hours until the reaction was complete. The resin was washed three times with 20 mL of DMF solution. After washing, proceed to the next step. Add 30 mL of a 20% Pip / DMF (v / v) solution, stir, and react for 30 minutes. Filter to remove the deprotection solution, then wash the resin six times with 30 mL of DMF solution, drain, and set aside.

[0163] Place 10 mmol of Fmoc-Leu-OH and 10 mmol of HOBt in a 100 mL beaker, cool to 5°C, add 15 mL of DMF solution and 1.5 mL of DIC, and allow to react for 15 minutes. The solution in the 100 mL beaker was then added to the 100 mL solid-phase synthesis reactor described above and stirred for 1.5 hours. The reaction was complete. The resin was washed three times with 20 mL of DMF solution. After washing, proceed to the next step. Add 30 mL of a 20% Pip / DMF (v / v) solution, stir, and react for 30 minutes. Filter to remove the deprotection solution, then wash the resin six times with 30 mL of DMF solution, drain, and set aside.

[0164] Place 10 mmol of Fmoc-Arg(Pbf)-OH and 10 mmol of HOBt in a 100 mL beaker, cool to 5°C, add 15 mL of DMF solution and 1.5 mL of DIC, and allow to react for 15 minutes. The solution in the 100 mL beaker was then added to the 100 mL solid-phase synthesis reactor described above and stirred for 1.5 hours until the reaction was complete. The resin was washed three times with 20 mL of DMF solution. After washing, proceed to the next step. Add 40 mL of a 20% Pip / DMF (v / v) solution, stir, and react for 30 minutes. Filter to remove the deprotection solution, then wash the resin six times with 40 mL of DMF solution, drain, and set aside.

[0165] Take 10mmol Fmoc-Phe-OH and 10mmol HOBt in a 100mL beaker, cool to 5℃, add 15mL of DMF solution and 1.5mL of DIC, let it stand for 15min, and add the solution in the 100mL beaker to the above 100mL solid phase synthesis reactor, stir and react for 1.5h, and the reaction is complete. Wash the resin with DMF solution 3 times, 20mL each time. After washing, proceed to the next step of the reaction. Add 40mL of 20% Pip / DMF (v / v) solution, stir and react for 30min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 40mL each time, and drain for use. 15 mmol of Fmoc-Trp(Boc)-OH and 1 mmol of HOBt were placed in a 100 mL beaker and cooled to 5°C. 15 mL of DMF solution and 2.3 mL of DIC were added and allowed to react for 15 minutes. The solution in the 100 mL beaker was then added to the 100 mL solid-phase synthesis reactor described above and stirred for 1.5 hours until the reaction was complete. The resin was washed three times with 30 mL of DMF solution. After washing, the resin was proceeded to the next step. 40 mL of a 20% Pip / DMF (v / v) solution was added and stirred for 30 minutes. The deprotection solution was removed by filtration and then washed six times with 40 mL of DMF solution. The mixture was then dried and set aside. The resin was then washed twice with 40 mL of methanol, twice with 40 mL of DCM solution, and twice with 40 mL of methanol. Drying under vacuum gave a peptide resin of H-AA1-AA2-AA3-AA4-AA5-AA6-Linker-AM resin, wherein AA1 is Trp(Boc); AA2 is Phe; AA3 is Arg; AA4 is D-Leu; AA5 is Ala; and AA6 is His(Trt).

[0166] The peptide resin was cleaved with 110 mL of TFA / thioanisole / phenol / H2O / EDT (the mass ratio of TFA, thioanisole, phenol, H2O, and EDT was 87.5:5:2.5:2.5:2.5) for 2.5 h. The cleavage solution was added to 1000 mL of ether (5°C) to precipitate a white solid, which was centrifuged and dried in vacuo to obtain H-Trp-Phe-Arg-Leu-Ala-His-NH2. Its mass spectrometry and HPLC characterization results are shown in Figure 2. Figure 86 and Figure 87 shown.

[0167] A synthetic route for a hyaluronic acid-modified beauty peptide is: ; Wherein, -COOX is -COONa; The preparation method of hyaluronic acid-modified cosmetic peptide comprises the following steps: Weigh 1g of H-Trp-Phe-Arg-Leu-Ala-His-NH2.TFA, add 10mL of DMSO and stir to dissolve, add 1.28g of DIEA, stir for 5 minutes, then add 3.75g of hyaluronic acid, control the temperature in a water bath at 45°C, stir and react overnight, take a sample and detect by LC-MS, if the reaction is basically complete, add 8mL of acetic acid, keep the temperature constant for rearrangement reaction for 2h, take a sample and detect by LC-MS, if the reaction is basically complete; then perform reverse phase chromatography purification, purification conditions: Dissolution: dilute 0.2 g of crude product with 100 mL of H2O; Packing: 21.2*250mm, 10-120, C18; Flow rate: 10mL / min; Wavelength: 220nm; Mobile phase: A: 1% HAc; B: ACN; Balance: A:B=95:5, balance 10 min, flow rate: 10 mL / min; Sample loading: flow rate: 10 mL / min; Elution: 12-32%B 60 min; Column cleaning: 80% ACN cleaning to baseline equilibrium; Qualified products were collected to obtain three structures of hyaluronic acid-modified beauty peptides, as shown below: B2, its mass spectrometry and HPLC characterization results are as follows Figure 88 and Figure 89 As shown; B4, its mass spectrometry and HPLC characterization results are as follows Figure 90 and Figure 91 As shown; B6, its mass spectrometry and HPLC characterization results are shown in Figure 92 and Figure 93 shown.

[0168] Example 29: A method for synthesizing H-Arg-Arg-Gln-D-Met-Glu-Glu-NH2, comprising: Place 5 mmol AM resin in a 100 mL solid-phase synthesis reactor, add 30 mL of 20% Pip / DMF (v / v) solution, stir and react for 30 min, filter to remove the deprotection solution, then wash with DMF solution 6 times, 30 mL each time, and drain to set aside.

[0169] Place 10 mmol of Fmoc-Linker and 10 mmol of HOBt in a 100 mL beaker, cool to 5°C, add 15 mL of DMF solution and 1.5 mL of DIC, and allow to react for 15 minutes. The solution in the 100 mL beaker was then added to the 100 mL solid-phase synthesis reactor described above and stirred for 1.5 hours until the reaction was complete. The resin was washed three times with 20 mL of DMF solution. After washing, proceed to the next step: add 30 mL of 20% Pip / DMF (v / v) solution, stir for 30 minutes, filter to remove the deprotection solution, and then wash six times with 30 mL of DMF solution before draining.

[0170] Take 10mmol Fmoc-Glu(otBu)-OH and 10mmol HOBt in a 100mL beaker, cool to 5℃, add 15mL of DMF solution and 1.5mL of DIC, let it stand for 15min, and add the solution in the 100mL beaker to the above 100mL solid-phase synthesis reactor. Stir and react for 1.5h until the reaction is complete. Wash the resin with DMF solution 3 times, 20mL each time. After washing, proceed to the next step. Add 40mL of 20% Pip / DMF (v / v) solution, stir and react for 30min, filter to remove the deprotection solution, then wash with DMF solution 6 times, 40mL each time, and drain for use; Take 10mmol Fmoc-Glu(otBu)-OH and 10mmol HOBt in a 100mL beaker, cool to 5℃, add 15mL of DMF solution and 1.5mL of DIC, let it stand for 15min, and add the solution in the 100mL beaker to the above 100mL solid-phase synthesis reactor. Stir and react for 1.5h until the reaction is complete. Wash the resin with DMF solution 3 times, 20mL each time. After washing, proceed to the next step. Add 40mL of 20% Pip / DMF (v / v) solution, stir and react for 30min, filter to remove the deprotection solution, then wash with DMF solution 6 times, 40mL each time, and drain for use; Place 10 mmol of Fmoc-D-Met-OH and 10 mmol of HOBt in a 100 mL beaker, cool to 5°C, add 15 mL of DMF solution and 1.5 mL of DIC, and allow to react for 15 minutes. The solution in the 100 mL beaker was then added to the 100 mL solid-phase synthesis reactor described above and stirred for 1.5 hours until the reaction was complete. Wash the resin three times with 20 mL of DMF solution. After washing, proceed to the next step. Add 40 mL of a 20% Pip / DMF (v / v) solution, stir, and react for 30 minutes. Filter to remove the deprotection solution, then wash the resin six times with 40 mL of DMF solution, drain, and set aside.

[0171] Place 10 mmol of Fmoc-Gln(Trt)-OH and 10 mmol of HOBt in a 100 mL beaker, cool to 5°C, add 15 mL of DMF solution and 1.5 mL of DIC, and allow to react for 15 minutes. The solution in the 100 mL beaker was then added to the 100 mL solid-phase synthesis reactor described above and stirred for 1.5 hours. The reaction was complete. The resin was washed three times with 20 mL of DMF solution. After washing, the resin was ready for the next step. 30 mL of a 20% Pip / DMF (v / v) solution was added and stirred for 30 minutes. The deprotection solution was removed by filtration, and the resin was then washed six times with 30 mL of DMF solution. The resin was then dried and used.

[0172] Place 10 mmol of Fmoc-Arg(Pbf)-OH and 10 mmol of HOBt in a 100 mL beaker, cool to 5°C, add 15 mL of DMF solution and 1.5 mL of DIC, and allow to react for 15 minutes. The solution in the 100 mL beaker was then added to the 100 mL solid-phase synthesis reactor described above and stirred for 1.5 hours until the reaction was complete. The resin was washed three times with 20 mL of DMF solution. After washing, proceed to the next step. Add 30 mL of a 20% Pip / DMF (v / v) solution, stir, and react for 30 minutes. Filter to remove the deprotection solution, then wash the resin six times with 30 mL of DMF solution, drain, and set aside.

[0173] 15 mmol of Fmoc-Arg(Pbf)-OH and 15 mmol of HOBt were placed in a 100 mL beaker and cooled to 5°C. 20 mL of DMF solution and 2.3 mL of DIC were added and allowed to react for 15 minutes. The solution in the 100 mL beaker was then added to the 100 mL solid-phase synthesis reactor described above and stirred for 1.5 hours until the reaction was complete. The resin was washed three times with 20 mL of DMF solution. After washing, the resin was proceeded to the next step. 40 mL of a 20% Pip / DMF (v / v) solution was added and stirred for 30 minutes. The deprotection solution was removed by filtration and then washed six times with 40 mL of DMF solution. The mixture was then dried and set aside. The resin was then washed twice with 40 mL of methanol, twice with 40 mL of DCM solution, and twice with 40 mL of methanol. Vacuum drying gave a peptide resin of H-AA1-AA2-AA3-AA4-AA5-AA6-Linker-AM resin, wherein AA1 is Arg(Pbf); AA2 is Arg(Pbf); AA3 is Gln(Trt); AA4 is D-Met; AA5 is Glu(otBu); and AA6 is Glu(otBu).

[0174] The peptide resin was cleaved with 110 mL of TFA / thioanisole / phenol / H2O / EDT (the mass ratio of TFA, thioanisole, phenol, H2O and EDT was 87.5:5:2.5:2.5:2.5) for 2.5 h. The cleavage solution was added to 1000 mL of ether (5°C) to precipitate a white solid, which was centrifuged and dried in vacuo to obtain H-Arg-Arg-Gln-D-Met-Glu-Glu-NH2. Its mass spectrometry and HPLC characterization results are shown in Figure 2. Figure 94 and Figure 95 shown.

[0175] A synthetic route for a hyaluronic acid-modified beauty peptide is: ; Wherein, -COOX is -COONa; The preparation method of hyaluronic acid-modified cosmetic peptide comprises the following steps: Weigh 1g of H-Arg-Arg-Gln-D-Met-Glu-Glu-NH2.TFA, add 10mL of DMSO and stir to dissolve, add 1.25g of DIEA, stir for 5 minutes, then add 3.67g of hyaluronic acid, control the temperature in a water bath at 45°C, stir and react overnight, take a sample and detect by LC-MS, if the reaction is basically complete, add 8mL of acetic acid, keep the temperature constant for rearrangement reaction for 2h, take a sample and detect by LC-MS, if the reaction is basically complete, then perform reverse phase chromatography purification, purification conditions: Dissolution: dilute 0.2 g of crude product with 100 mL of H2O; Packing: 21.2*250mm, 10-120, C18; Flow rate: 10mL / min; Wavelength: 220nm; Mobile phase: A: 1% HAc; B: ACN; Balance: A:B=95:5, balance 10 min, flow rate: 10 mL / min; Sample loading: flow rate: 10 mL / min; Elution: 12-32%B 60 min; Column cleaning: 80% ACN cleaning to baseline equilibrium; Qualified products were collected to obtain three structures of hyaluronic acid-modified beauty peptides, as shown below: G2, its mass spectrometry and HPLC characterization results are as follows Figure 96 and Figure 97 As shown; G4, its mass spectrometry and HPLC characterization results are as follows Figure 98 and Figure 99 As shown; G6, its mass spectrometry and HPLC characterization results are shown in Figure 100 and Figure 101 shown.

[0176] Example 30: A method for synthesizing H-Arg-Arg-Gln-Met-Glu-Glu-NH2, comprising: Place 5 mmol AM resin in a 100 mL solid-phase synthesis reactor, add 30 mL of 20% Pip / DMF (v / v) solution, stir and react for 30 min, filter to remove the deprotection solution, then wash with DMF solution 6 times, 30 mL each time, and drain to set aside.

[0177] Place 10 mmol of Fmoc-Linker and 10 mmol of HOBt in a 100 mL beaker, cool to 5°C, add 15 mL of DMF solution and 1.5 mL of DIC, and allow to react for 15 minutes. The solution in the 100 mL beaker was then added to the 100 mL solid-phase synthesis reactor described above and stirred for 1.5 hours until the reaction was complete. The resin was washed three times with 20 mL of DMF solution. After washing, proceed to the next step: add 30 mL of 20% Pip / DMF (v / v) solution, stir for 30 minutes, filter to remove the deprotection solution, and then wash six times with 30 mL of DMF solution before draining.

[0178] Take 10mmol Fmoc-Glu(otBu)-OH and 10mmol HOBt in a 100mL beaker, cool to 5℃, add 15mL of DMF solution and 1.5mL of DIC, let it stand for 15min, and add the solution in the 100mL beaker to the above 100mL solid phase synthesis reactor, stir and react for 1.5h, and the reaction is complete. Wash the resin with DMF solution 3 times, 20mL each time. After washing, proceed to the next step of the reaction. Add 40mL of 20% Pip / DMF (v / v) solution, stir and react for 30min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 40mL each time, and drain and set aside. Take 10mmol Fmoc-Glu(otBu)-OH and 10mmol HOBt in a 100mL beaker, cool to 5℃, add 15mL of DMF solution and 1.5mL of DIC, let it stand for 15min, and add the solution in the 100mL beaker to the above 100mL solid phase synthesis reactor, stir and react for 1.5h, and the reaction is complete. Wash the resin with DMF solution 3 times, 20mL each time. After washing, proceed to the next step of the reaction. Add 40mL of 20% Pip / DMF (v / v) solution, stir and react for 30min, filter to remove the deprotection solution, and then wash with DMF solution 6 times, 40mL each time, and drain and set aside. Place 10 mmol of Fmoc-Met-OH and 10 mmol of HOBt in a 100 mL beaker, cool to 5°C, add 15 mL of DMF solution and 1.5 mL of DIC, and allow to react for 15 minutes. The solution in the 100 mL beaker was then added to the 100 mL solid-phase synthesis reactor described above and stirred for 1.5 hours. The reaction was complete. The resin was washed three times with 20 mL of DMF solution. After washing, proceed to the next step. Add 40 mL of a 20% Pip / DMF (v / v) solution and stir for 30 minutes. Filter to remove the deprotection solution, then wash the resin six times with 40 mL of DMF solution, draining thoroughly for later use.

[0179] Place 10 mmol of Fmoc-Gln(Trt)-OH and 10 mmol of HOBt in a 100 mL beaker, cool to 5°C, add 15 mL of DMF solution and 1.5 mL of DIC, and allow to react for 15 minutes. The solution in the 100 mL beaker was then added to the 100 mL solid-phase synthesis reactor described above and stirred for 1.5 hours. The reaction was complete. The resin was washed three times with 20 mL of DMF solution. After washing, the resin was ready for the next step. 30 mL of a 20% Pip / DMF (v / v) solution was added and stirred for 30 minutes. The deprotection solution was removed by filtration, and the resin was then washed six times with 30 mL of DMF solution. The resin was then dried and used.

[0180] Place 10 mmol of Fmoc-Arg(Pbf)-OH and 10 mmol of HOBt in a 100 mL beaker, cool to 5°C, add 15 mL of DMF solution and 10 mmol of DIC, and let the mixture react for 15 minutes. The mixture was then added to the 100 mL solid-phase synthesis reactor described above and stirred for 1.5 hours. The reaction was complete. The resin was washed three times with 20 mL of DMF solution. After washing, the resin was ready for the next step. 30 mL of a 20% Pip / DMF (v / v) solution was added and stirred for 30 minutes. The deprotection solution was removed by filtration, and the resin was then washed six times with 30 mL of DMF solution. The mixture was then dried and used.

[0181] 15 mmol of Fmoc-Arg(Pbf)-OH and 15 mmol of HOBt were placed in a 100 mL beaker and cooled to 5°C. 20 mL of DMF solution and 2.3 mL of DIC were added and allowed to react for 15 minutes. The solution in the 100 mL beaker was then added to the 100 mL solid-phase synthesis reactor described above and stirred for 1.5 hours until the reaction was complete. The resin was washed three times with 20 mL of DMF solution each time. After washing, the resin was processed for the next step. 40 mL of 20% Pip / DMF (v / v) solution was added and stirred for 30 minutes. The deprotection solution was removed by filtration and then washed six times with 40 mL of DMF solution. The mixture was then dried and set aside. The resin was then washed twice with 40 mL of methanol, twice with 40 mL of DCM solution, and twice with 40 mL of methanol. Drying under vacuum gave a peptide resin of H-AA1-AA2-AA3-AA4-AA5-AA6-Linker-AM resin, wherein AA1 is Arg(Pbf); AA2 is Arg(Pbf); AA3 is Gln(Trt); AA4 is Met; AA5 is Glu(otBu); and AA6 is Glu(otBu).

[0182] The peptide resin was cleaved with 110 mL of TFA / thioanisole / phenol / H2O / EDT (the mass ratio of TFA, thioanisole, phenol, H2O and EDT was 87.5:5:2.5:2.5:2.5) for 2.5 h. The cleavage solution was added to 1000 mL of ether (5°C) to precipitate a white solid, which was centrifuged and dried in vacuo to obtain H-Arg-Arg-Gln-Met-Glu-Glu-NH2. Its mass spectrometry and HPLC characterization results are shown in Figure 2. Figure 102 and Figure 103 shown.

[0183] A synthetic route for a hyaluronic acid-modified beauty peptide is: ; Wherein, -COOX is -COONa; The preparation method of hyaluronic acid-modified cosmetic peptide comprises the following steps: Weigh 1g of H-Arg-Arg-Gln-Met-Glu-Glu-NH2.TFA, add 10mL of DMSO and stir to dissolve, add 1.25g of DIEA, stir for 5 minutes, then add 3.67g of hyaluronic acid, control the temperature in a water bath at 45°C, stir and react overnight, take a sample and detect by LC-MS, if the reaction is basically complete, add 8mL of acetic acid, keep the temperature constant for rearrangement reaction for 2h, take a sample and detect by LC-MS, if the reaction is basically complete, then perform reverse phase chromatography purification, purification conditions: Dissolution: dilute 0.2 g of crude product with 100 mL of H2O; Packing: 21.2*250mm, 10-120, C18; Flow rate: 10mL / min; Wavelength: 220nm; Mobile phase: A: 1% HAc; B: ACN; Balance: A:B=95:5, balance 10 min, flow rate: 10 mL / min; Sample loading: flow rate: 10 mL / min; Elution: 12-32%B 60 min; Column cleaning: 80% ACN cleaning to baseline equilibrium; Qualified products were collected to obtain three structures of hyaluronic acid-modified beauty peptides, as shown below: F2, its mass spectrometry and HPLC characterization results are as follows Figure 104 and Figure 105 As shown; F4, its mass spectrometry and HPLC characterization results are as follows Figure 106 and Figure 107 As shown; F6, its mass spectrometry and HPLC characterization results are shown in Figure 108 and Figure 109 shown.

[0184] Example 31: A method for preparing a hyaluronic acid-modified cosmetic peptide comprises the following steps: Weigh 0.56 g H-Phe-Val-Ala-Pro-Phe-Pro-OH.TFA (prepared in Example 8), add 6 mL DMSO and stir to dissolve, add 0.75 g DIEA, then add 2.2 g hyaluronic acid (y=1), control the temperature in a water bath at 45°C, stir and react overnight, take a sample for LC-MS detection, and the reaction is basically complete, add 4 mL acetic acid, control the temperature in a water bath at 35°C, and carry out rearrangement reaction for 3 hours, take a sample for LC-MS detection, and the reaction is basically complete; then perform reverse chromatography purification to obtain the hyaluronic acid-modified beauty peptide.

[0185] Example 32: A method for preparing a hyaluronic acid-modified cosmetic peptide comprises the following steps: Weigh 0.56 g H-Phe-Val-Ala-Pro-Phe-Pro-OH.TFA (prepared in Example 8), add 6 mL DMSO and stir to dissolve, add 0.75 g DIEA, then add 2.2 g hyaluronic acid (y=2), control the temperature in a water bath at 45°C, stir and react overnight, take a sample for LC-MS detection, and the reaction is basically complete, add 4 mL acetic acid, control the temperature in a water bath at 35°C, and carry out rearrangement reaction for 3 hours, take a sample for LC-MS detection, and the reaction is basically complete; then perform reverse chromatography purification to obtain the hyaluronic acid-modified beauty peptide.

[0186] Example 33: A method for preparing a hyaluronic acid-modified cosmetic peptide comprises the following steps: Weigh 0.56 g H-Phe-Val-Ala-Pro-Phe-Pro-OH.TFA (prepared in Example 8), add 6 mL DMSO and stir to dissolve, add 0.75 g DIEA, then add 2.2 g hyaluronic acid (y=3), control the temperature in a water bath at 45°C, stir and react overnight, take a sample for LC-MS detection, and the reaction is basically complete, add 4 mL acetic acid, control the temperature in a water bath at 35°C, and carry out rearrangement reaction for 3 hours, take a sample for LC-MS detection, and the reaction is basically complete; then perform reverse chromatography purification to obtain the hyaluronic acid-modified beauty peptide.

[0187] Test Example 1: 1. Moisturizing performance test AQP3 content is tested as follows: (1) Cell seeding: Cells were seeded into 24-well plates and incubated overnight in an incubator (37°C, 5% CO2); (2) Solution preparation: Prepare the working solution of the test substance according to the experimental design (as shown in Table 1); Table 1 AQP3 experimental design table

[0188] (3) Adding the test substance: After culturing in an incubator (37°C, 5% CO2) for 24 h, add the test substance according to the table and continue culturing for another 24 h; (4) Sample collection: discard the supernatant and rinse the cells three times with PBS; (5) Immunofluorescence staining: a. Add methanol to fix the cells, rinse three times with PBS, and then add 1 mL of BSA to each well to block for 1 hour; b. Discard the blocking solution, add primary antibody to each well, and place in a 4°C refrigerator overnight. Discard the primary antibody and rinse three times with PBS; c. Add secondary antibody to each well and allow to react for 2 hours; then discard the secondary antibody and rinse three times with PBS; d. Add DAPI to each well for nuclear staining for 10 min, discard the DAPI, rinse three times with PBS, and then take pictures using a fluorescence microscope; (6) Result analysis: AQP3 fluorescence intensity was quantitatively analyzed using Image Pro Plus software.

[0189] HA content test is as follows: (1) Cell seeding: Cells were seeded into 24-well plates and incubated overnight in an incubator (37°C, 5% CO2); (2) Solution preparation: Prepare the working solution of the test substance according to the experimental design (Table 2); Table 2 HA experimental design table

[0190] (3) Adding the test substance: After culturing in an incubator (37°C, 5% CO2) for 24 hours, add the test substance according to the table and continue culturing for another 24 hours; (4) Sample collection: Collect the supernatant and determine the HA content using an ELISA kit.

[0191] Result analysis: Table 3-1 AQP3 test results

[0192] Analysis of the data in Table 3-1 demonstrates that the hyaluronic acid-modified cosmetic peptides prepared in the Examples of the present invention exhibit excellent moisturizing properties and are capable of effectively increasing AQP3 content. Specifically, the hyaluronic acid-modified cosmetic peptides D4 and D6 prepared in Example 4 exhibited excellent moisturizing efficacy at concentrations of 0.063 mg / mL, 0.125 mg / mL, and 0.25 mg / mL, respectively. Furthermore, when the three hyaluronic acid-modified cosmetic peptides (D2, D4, and D6) were mixed at a specific mass ratio, they also exhibited excellent moisturizing efficacy at concentrations of 0.063 mg / mL, 0.125 mg / mL, and 0.25 mg / mL. The three hyaluronic acid-modified cosmetic peptides prepared in Example 27 of the present invention, when mixed at a certain mass ratio, exhibited excellent moisturizing efficacy at concentrations of 0.063 mg / mL, 0.125 mg / mL, and 0.25 mg / mL. The three hyaluronic acid-modified cosmetic peptides prepared in Example 28 of the present invention, when mixed at a certain mass ratio, exhibited excellent moisturizing efficacy at concentrations of 0.063 mg / mL, 0.125 mg / mL, and 0.25 mg / mL. The three hyaluronic acid-modified cosmetic peptides prepared in Example 29 of the present invention, when mixed at a certain mass ratio, exhibited excellent moisturizing efficacy at concentrations of 0.063 mg / mL, 0.125 mg / mL, and 0.25 mg / mL. The three hyaluronic acid-modified cosmetic peptides (F2, F4, and F6) prepared in Example 30 of the present invention, when mixed at a certain mass ratio, exhibited excellent moisturizing efficacy at concentrations of 0.063 mg / mL, 0.125 mg / mL, and 0.25 mg / mL. The hyaluronic acid-modified cosmetic peptides A4 and A6 prepared in Example 14 of the present invention exhibited excellent moisturizing efficacy at concentrations of 0.063 mg / mL, 0.125 mg / mL, and 0.25 mg / mL. Furthermore, the three hyaluronic acid-modified cosmetic peptides (A2, A4, and A6), when mixed at a certain mass ratio, also exhibited excellent moisturizing efficacy at concentrations of 0.063 mg / mL, 0.125 mg / mL, and 0.25 mg / mL.

[0193] Table 3-2 AQP3 test results

[0194] Analysis of the data in Table 3-2 demonstrates that the hyaluronic acid-modified cosmetic peptides prepared in the Examples of the present invention exhibit excellent AQP3-boosting effects, particularly enhancing its content. Specifically, the hyaluronic acid-modified cosmetic peptides prepared in Examples 1-2, 5-7, and 10-13 of the present invention exhibited excellent moisturizing effects at concentrations of 0.063 mg / mL, 0.125 mg / mL, and 0.25 mg / mL. In addition, the three structures (D2, D4, and D6) of hyaluronic acid-modified beauty peptides prepared in Example 4 of the present invention, after being mixed in a certain mass ratio, have excellent moisturizing effects at concentrations of 0.063 mg / mL, 0.125 mg / mL, and 0.25 mg / mL; the three structures (H2, H4, and H6) of hyaluronic acid-modified beauty peptides prepared in Example 8 of the present invention, after being mixed in a certain mass ratio, have excellent moisturizing effects at concentrations of 0.063 mg / mL, 0.125 mg / mL, and 0.25 mg / mL; the three structures (L2, L4, and L6) of hyaluronic acid-modified beauty peptides prepared in Example 9 of the present invention, after being mixed in a certain mass ratio, have excellent moisturizing effects at concentrations of 0.063 mg / mL, 0.125 mg / mL, and 0.25 mg / mL.

[0195] Table 4 HA test results

[0196] Analysis of the data in Table 4 demonstrates that the hyaluronic acid-modified cosmetic peptides prepared in Examples 2 and 8 of the present invention exhibit excellent hydration-enhancing effects, particularly enhancing HA content. Specifically, the hyaluronic acid-modified cosmetic peptide prepared in Example 2 exhibits excellent moisturizing efficacy at concentrations of 0.063 mg / mL, 0.125 mg / mL, and 0.25 mg / mL. Furthermore, the three hyaluronic acid-modified cosmetic peptides (H2, H4, and H6) prepared in Example 8, when mixed at a specific mass ratio, exhibit excellent moisturizing efficacy at concentrations of 0.063 mg / mL, 0.125 mg / mL, and 0.25 mg / mL.

[0197] 2. Anti-wrinkle and firming performance measurement Type I collagen and MMP-1 content were tested as follows: (1) Cell seeding: Cells were seeded into 24-well plates and incubated overnight in an incubator (37°C, 5% CO2); (2) Solution preparation: Prepare the working solution of the test substance according to the experimental design (Table 5); Table 5 Experimental design table

[0198] (3) UVA radiation: After 24 hours of culture, the negative control group, positive control group and sample group received a total dose of 9 J / cm 2 UVA radiation, while the blank control group was placed in the same environment (UVA radiation dose of 0J / cm 2 ); (4) Adding the test substance: According to the experimental design, after irradiation, the test substance was added to each group. 1 mL of cell culture medium was added to each well of the blank control group and the negative control group; 1 mL of cell culture medium containing vitamin C and vitamin E was added to each well of the positive control group; 1 mL of culture medium containing the corresponding concentration of the test substance was added to each well of the sample group; after the addition of the test substance, the 24-well plate was placed in an incubator (37°C, 5% CO2) and cultured for 24 h; (5) Collect the supernatant and determine the content of type I collagen and MMP-1; (6) Result analysis: The comparison between the groups was performed using t-test statistical analysis, and all statistical analyses were two-tailed.

[0199] Table 6 MMP-1 test results

[0200] Table 7-1 Type I collagen test results

[0201] Analysis of the data in Tables 6 and 7-1 demonstrates that the hyaluronic acid-modified cosmetic peptides prepared in the Examples of the present invention exhibit excellent inhibitory effects on MMP-1 and excellent promoting effects on Collagen I. Specifically, the hyaluronic acid-modified cosmetic peptides D4 and D6 prepared in Example 4 of the present invention exhibited excellent anti-wrinkle and firming effects at concentrations of 0.063 mg / mL, 0.125 mg / mL, and 0.25 mg / mL, respectively. Furthermore, when the three hyaluronic acid-modified cosmetic peptides (D2, D4, and D6) were mixed at a specific mass ratio, they also exhibited excellent anti-wrinkle and firming effects at concentrations of 0.063 mg / mL, 0.125 mg / mL, and 0.25 mg / mL. The three hyaluronic acid-modified cosmetic peptides prepared in Example 27 of the present invention, when mixed at a certain mass ratio, exhibited excellent anti-wrinkle and firming effects at concentrations of 0.063 mg / mL, 0.125 mg / mL, and 0.25 mg / mL. The three hyaluronic acid-modified cosmetic peptides prepared in Example 28 of the present invention, when mixed at a certain mass ratio, exhibited excellent anti-wrinkle and firming effects at concentrations of 0.063 mg / mL, 0.125 mg / mL, and 0.25 mg / mL. The three hyaluronic acid-modified cosmetic peptides prepared in Example 29 of the present invention, when mixed at a certain mass ratio, exhibited excellent anti-wrinkle and firming effects at concentrations of 0.063 mg / mL, 0.125 mg / mL, and 0.25 mg / mL. The three hyaluronic acid-modified cosmetic peptides (F2, F4, and F6) prepared in Example 30 of the present invention, when mixed at a certain mass ratio, exhibited excellent anti-wrinkle and firming efficacy at concentrations of 0.063 mg / mL, 0.125 mg / mL, and 0.25 mg / mL. The hyaluronic acid-modified cosmetic peptides A4 and A6 prepared in Example 14 of the present invention exhibited excellent anti-wrinkle and firming efficacy at concentrations of 0.063 mg / mL, 0.125 mg / mL, and 0.25 mg / mL. Furthermore, the three hyaluronic acid-modified cosmetic peptides (A2, A4, and A6), when mixed at a certain mass ratio, also exhibited excellent anti-wrinkle and firming efficacy at concentrations of 0.063 mg / mL, 0.125 mg / mL, and 0.25 mg / mL.

[0202] Table 7-2 Type I collagen test results

[0203] From the data analysis in Table 7-2, it can be seen that the hyaluronic acid-modified cosmetic peptides prepared in Examples 1-2, 5-7 and 10-13 of the present invention have excellent anti-wrinkle and firming effects at concentrations of 0.063 mg / mL, 0.125 mg / mL and 0.25 mg / mL. In addition, the three hyaluronic acid-modified beauty peptides prepared in Example 4 of the present invention, after being mixed in a certain mass ratio, have excellent anti-wrinkle and firming effects at concentrations of 0.063 mg / mL, 0.125 mg / mL and 0.25 mg / mL; the three hyaluronic acid-modified beauty peptides prepared in Example 8 of the present invention, after being mixed in a certain mass ratio, have excellent anti-wrinkle and firming effects at concentrations of 0.063 mg / mL, 0.125 mg / mL and 0.25 mg / mL; the three hyaluronic acid-modified beauty peptides prepared in Example 9 of the present invention, after being mixed in a certain mass ratio, have excellent anti-wrinkle and firming effects at concentrations of 0.063 mg / mL, 0.125 mg / mL and 0.25 mg / mL.

[0204] 3. Soothing performance test IL-6 content test (1) Cell seeding: Cells were seeded into 24-well plates and incubated overnight in an incubator (37°C, 5% CO2); (2) Solution preparation: Prepare the working solution of the test substance according to the experimental design (as shown in Table 8); Table 8 IL-6 synthesis experimental design table

[0205] (3) Adding the test substance: According to the experimental grouping, when the cell plating rate in the 24-well plate reaches 40%-60%, the test substance is added in groups, with three replicate wells in each group. The 24-well plate is placed in an incubator (37°C, 5% CO2) and incubated for 24 h; (4) Detection: After 24 hours of culture, the supernatant was collected and the IL-6 content was determined using an ELISA kit.

[0206] Table 9 Soothing performance test results

[0207] From the data analysis in Table 9, it can be seen that the hyaluronic acid-modified beauty peptide prepared in Example 2 of the present invention has a soothing effect at concentrations of 0.063 mg / mL, 0.125 mg / mL and 0.25 mg / mL; the hyaluronic acid-modified beauty peptide prepared in Example 5 of the present invention has a soothing effect at a concentration of 0.063 mg / mL; the hyaluronic acid-modified beauty peptide prepared in Example 6 of the present invention has a soothing effect at concentrations of 0.063 mg / mL, 0.125 mg / mL and 0.25 mg / mL; the hyaluronic acid-modified beauty peptide prepared in Example 7 of the present invention has a soothing effect at a concentration of 0.125 mg / mL; the three structures of the hyaluronic acid-modified beauty peptides prepared in Example 9 of the present invention have a soothing effect. The hyaluronic acid-modified beauty peptide prepared in Example 10 of the present invention has a soothing effect at a concentration of 0.063 mg / mL; the hyaluronic acid-modified beauty peptide prepared in Example 13 of the present invention has a soothing effect at a concentration of 0.125 mg / mL; the hyaluronic acid-modified beauty peptide prepared in Example 11 of the present invention has a soothing effect at concentrations of 0.063 mg / mL and 0.125 mg / mL; the hyaluronic acid-modified beauty peptide prepared in Example 12 of the present invention has a soothing effect at concentrations of 0.063 mg / mL and 0.25 mg / mL.

[0208] 4. Whitening performance test Melanin content test is as follows: Table 10 Melanin content experimental design table

[0209] Cells in the logarithmic growth phase were collected and inoculated into 24-well plates. After culturing in an incubator (37°C, 5% CO2) for 24 h, the test substances were added according to the table based on the cytotoxicity results. Untreated cells were used as blank controls. Three parallels were set up for each group.

[0210] After adding the drug, continue culturing in an incubator (37°C, 5% CO2) for 24 hours, discard the supernatant, add 0.5 mL of 1M NaOH containing 10% DMSO, and incubate at 80°C for 1 hour. Use 1M NaOH containing 10% DMSO as the solvent control, read the absorbance value on a microplate reader, and calculate the relative inhibition rate of cytomelanin.

[0211]

[0212] Table 11 Whitening performance test results

[0213] From the data analysis in Table 11, it can be seen that the hyaluronic acid-modified cosmetic peptide prepared in Example 7 of the present invention has excellent whitening efficacy at concentrations of 0.063 mg / mL, 0.125 mg / mL and 0.25 mg / mL; and the whitening effects at concentrations of 0.125 mg / mL and 0.25 mg / mL are comparable to or higher than that of the positive control group.

[0214] 5. Anti-glycation performance test Prepare a mixed solution containing bovine serum albumin and glucose in PBS, filter through a 0.22 μm filter membrane, and use it as the 2× saccharification reaction solution. Prepare each reaction system according to the table.

[0215] Table 12 AGEs removal test reaction system

[0216] After mixing thoroughly, incubate at 55°C for 4 days. PBS was used instead of the sample as a negative control, aminoguanidine hydrochloride (100 mg / mL) as a positive control, and PBS was used instead of the glycosylation reaction solution as a control system. After the reaction, the incubation solution was cooled to room temperature and centrifuged at 2000 rpm for 5 minutes. The supernatant was filtered through a 0.22 μm filter membrane, and 200 μL of the reaction solution was added to a 96-well plate. Fluorescence microplate reader was used to detect the AGEs inhibition rate under the conditions of excitation wavelength of 320 nm and emission wavelength of 460 nm.

[0217]

[0218] Where: A-fluorescence intensity of the glycation system after adding the test substance; B-fluorescence intensity of PBS solution with added test substance; C-fluorescence intensity of the glycation system without adding the test substance; D- Fluorescence intensity of PBS solution without adding test substance.

[0219] Table 13 Anti-glycation performance test results

[0220] From the data analysis in Table 13, it can be seen that the hyaluronic acid-modified cosmetic peptide prepared in Example 11 of the present invention has a scavenging effect on advanced glycation end products at concentrations of 0.0625 mg / mL and 0.25 mg / mL.

[0221] 6. Antioxidant performance test ROS content was tested as follows: (1) Cell seeding: Cells were seeded into 24-well plates and incubated overnight in an incubator (37°C, 5% CO2); (2) Liquid preparation: Prepare the working solution of the test substance according to the experimental design (Table 13).

[0222] Table 14 ROS experimental design table

[0223] (3) Adding the test substance: After culturing in an incubator (37°C, 5% CO2) for 24 hours, add the test substance according to the table and continue culturing for another 24 hours; (4) Modeling: Washed with PBS twice, untreated cells were used as blank control, and the other groups were stimulated with UVB according to the conditions in the table, with the VC+VE group as the positive control; (5) ROS content detection: DCFH-DA probe stock solution was diluted with serum-free culture medium. 500 μL of the diluted DCFH-DA probe was added to each well and incubated in a 37°C cell culture incubator. After 30 min, the cells were washed three times with serum-free DMEM culture medium to fully remove the probe that did not enter the cells. The cells were observed and photographed using a fluorescence microscope at an excitation wavelength of 488 nm.

[0224] (6) Result analysis: Image Pro Plus software was used to quantitatively analyze the ROS fluorescence intensity.

[0225] Table 15 Antioxidant performance test results

[0226] From the data analysis in Table 15, it can be seen that the hyaluronic acid-modified cosmetic peptide prepared in Example 11 of the present invention has better antioxidant capacity at concentrations of 0.125 mg / mL and 0.25 mg / mL, and its antioxidant capacity level at a concentration of 0.25 mg / mL is comparable to that of the positive control.

[0227] The conventional techniques in the above embodiments are prior arts known to those skilled in the art, and thus will not be described in detail here.

[0228] The above description is merely a specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any modifications or substitutions that can be easily conceived by a person skilled in the art within the technical scope disclosed in the present invention should be included in the scope of protection of the present invention. Therefore, the scope of protection of the present invention should be based on the scope of protection of the claims.

Claims

1. A hyaluronic acid-modified beauty peptide, wherein the structural formula of the hyaluronic acid-modified beauty peptide is shown in formula (I): MC (I); in, M represents sodium hyaluronate, and its structure is shown in formula (II): (II), where y is a natural number ≥ 1; C represents a beauty peptide, which includes a polypeptide or a derivative thereof having beauty and / or skin care effects, and the polypeptide includes a heptapeptide; Among them, the M terminal glucuronic acid links the amino group in the X structure.

2. The hyaluronic acid-modified cosmetic peptide according to claim 1, characterized in that: The compound represented by formula (I) includes the structure represented by formula (III): (III); in, Said n is a natural number; The R is the remaining part of the cosmetic peptide structure after removing the reactive amino group; The cosmetic peptides include polypeptides or derivatives thereof having cosmetic and / or skin care effects, and the polypeptides include heptapeptides.

3. The hyaluronic acid-modified cosmetic peptide according to claim 1 or 2, characterized in that: The heptapeptide or its derivatives include a heptapeptide having an amino acid sequence of H-Glu-Glu-Met-Gln-Arg-Arg-Ala-OH or H-Glu-Glu-Met-Gln-Arg-Arg-Ala-NH2.

4. The method for preparing the hyaluronic acid-modified cosmetic peptide according to claim 1 or 2, comprising: Sodium hyaluronate is used to react with beauty peptides to produce hyaluronic acid-modified beauty peptides.

5. The method for preparing the hyaluronic acid-modified cosmetic peptide according to claim 4, characterized in that: The molar ratio of the cosmetic peptide to sodium hyaluronate is 1:2-8.

6. Use of the hyaluronic acid-modified beauty peptide obtained by the preparation method according to claim 4 in the preparation of cosmetics and / or skin care products.

7. Use of the hyaluronic acid-modified beauty peptide obtained by the preparation method of claim 4 in enhancing the moisturizing, firming, anti-wrinkle or anti-aging properties of cosmetics and / or skin care products.

8. Use of the hyaluronic acid-modified beauty peptide obtained by the preparation method according to claim 4 in enhancing the soothing, antioxidant or whitening properties of cosmetics and / or skin care products.

9. A cosmetic comprising the hyaluronic acid-modified cosmetic peptide according to claim 1.

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