An acetylhyaluronic acid oligopeptide and its preparation and application methods

Abstract

This invention relates to an acetyl hyaluronic acid oligopeptide and its preparation and application methods, solving the technical problems of existing technologies such as the instability of the structure of acetyl hyaluronic acid and oligopeptides, the need for separate addition of acetyl hyaluronic acid and oligopeptides in functional cosmetics, easy enzymatic hydrolysis, poor chemical stability, susceptibility to environmental influences leading to inactivation, and short duration of action. This invention provides a method for preparing an anti-wrinkle and anti-aging acetyl hyaluronic acid oligopeptide, comprising the following steps: linking acetyl hyaluronic acid and oligopeptide via covalent bonds: the ε-NH2 of the Lys side chain in the oligopeptide sequence and the -COOH of acetyl hyaluronic acid are linked through an amide condensation reaction to form an amide bond, thereby generating the acetyl hyaluronic acid oligopeptide; and simultaneously providing its application method. This invention has wide applications in the fields of functional cosmetics technology and plastic surgery products technology.

Description

[0001] This case is a divisional application based on Chinese Patent Application No. 2019109634683, filed on October 11, 2019, entitled "An Acetyl Hyaluronic Acid Oligopeptide and Its Preparation and Application Method". Technical Field

[0002] This invention belongs to the field of functional cosmetics technology, and specifically relates to an acetyl hyaluronic acid oligopeptide and its preparation and application methods. Background Technology

[0003] The development of functional cosmetics has become a major theme in the cosmetics industry. The added bioactive ingredients play a crucial role in regulating cell growth and metabolism. For example, peptides and polysaccharides can effectively address skin damage and promote skin repair. Peptides possess unique biological activity, playing a vital regulatory role in the body's growth, development, and metabolism. Their special physiological activity plays a crucial regulatory role in skin aging and care, and they are widely used in beauty and skincare products. Beauty peptides are mainly small molecule oligopeptides composed of two to ten amino acids. They can promote cell growth, differentiation, reconstruction, and repair, fundamentally improving and repairing skin damage. They have significant effects such as wrinkle reduction, anti-aging, whitening, and spot removal, and have become an important component of functional cosmetics with extremely high safety. For example, tripeptide-1 can stimulate the production of collagen, elastin, fibronectin, and laminin, restoring skin elasticity and firmness. It has anti-inflammatory, antioxidant, wrinkle-reducing, and anti-aging effects and can be used in the production of hair and skincare products. SIKVAV oligopeptides and IKVAV oligopeptides can promote cell adhesion, improve skin flexibility, and prevent skin aging. Hyaluronic acid (HA) is a glycosaminoglycan with exceptional water-retention properties. It is currently considered the best moisturizing substance found in nature, capable of retaining up to 1000 times its own weight in water. A 1% solution can form a gel, preventing moisture loss through the epidermis. It is considered an ideal natural moisturizing factor and also an excellent transdermal absorption enhancer. Hyaluronic acid's unique molecular structure and physicochemical properties provide various important physiological functions in the body, including preventing and repairing skin damage and promoting wound healing. It is widely used in the production of beauty and skincare products. Small-molecule hyaluronic acid can penetrate deep into the dermis, scavenging free radicals, improving skin nutrition and metabolism, promoting epidermal cell proliferation and differentiation, increasing skin elasticity, reducing wrinkles, and preventing aging. Large-molecule hyaluronic acid can form a protective film on the skin's surface, locking in moisture and blocking damage caused by adverse environmental factors. Acetylated hyaluronic acid (AcHA) is an acetylated derivative of hyaluronic acid. It is a novel and highly effective skin-softening factor with both hydrophilic and lipophilic properties. It can significantly soften the stratum corneum and has a strong affinity for the skin. Its water-retention capacity is more than twice that of hyaluronic acid, and its skin-care effects are superior. Functional cosmetics containing oligopeptides or hyaluronic acid are favored by consumers due to their safety, stability, easy absorption, and excellent effects. Furthermore, functional cosmetics have high added value and a large market potential, significantly increasing the profits of cosmetic manufacturers. As a result, more and more companies and research institutions are investing in the research and development of functional cosmetics.

[0004] Patent CN106109296B discloses a moisturizing essence water containing sodium hyaluronate and acetylated sodium hyaluronate; CN104740643A discloses a stable hyaluronic acid solution loaded with bioactive proteins or peptides, requiring the separate addition of hyaluronic acid and bioactive proteins or peptides; CN10

[0005] Patent 9157471A discloses a skincare composition with repairing and moisturizing functions, containing hyaluronic acid and oligopeptides respectively; CN108904317A discloses a method for combining anti-wrinkle peptides. However, these patents only add hyaluronic acid and peptides alone or in combination to the product, without mentioning hyaluronic acid oligopeptides that simultaneously possess the functions of two active ingredients, or their development and application in cosmetic compositions. A single active ingredient has limited effects on cells; using two active ingredients in combination, or using a component that simultaneously possesses the functions of two active ingredients, obviously achieves better results. However, human hyaluronidase and protease can hydrolyze hyaluronic acid and oligopeptides. Therefore, hyaluronic acid and oligopeptides absorbed through the skin and on the skin surface are easily deactivated by enzymatic degradation. Furthermore, due to the unstable structure of peptides, they are easily denatured and deactivated by adverse external environmental factors, resulting in short-lived activity. Therefore, it is necessary to select appropriate dosage forms and protective agents based on the characteristics of the ingredients to improve stability and prolong their efficacy. Summary of the Invention

[0006] The purpose of this invention is to overcome the shortcomings of the prior art and address the instability of the structures of acetyl hyaluronic acid and oligopeptides. It provides an acetyl hyaluronic acid oligopeptide that combines the functional activities of both acetyl hyaluronic acid and oligopeptides, is chemically stable, not easily affected by the environment, and has significant anti-wrinkle and anti-aging effects with a long duration of action, along with its preparation and application methods.

[0007] The technical solution adopted in this invention is as follows:

[0008] An anti-wrinkle and anti-aging acetyl hyaluronic acid oligopeptide, the acetyl hyaluronic acid oligopeptide having the structures of formulas V, VI and VII respectively:

[0009]

[0010] A method for preparing an anti-wrinkle and anti-aging acetyl hyaluronic acid oligopeptide includes the following steps: linking acetyl hyaluronic acid and oligopeptide through covalent bonds: the ε-NH2 of the Lys side chain in the oligopeptide sequence and the -COOH of acetyl hyaluronic acid are linked through an amide condensation reaction to generate an amide bond, thereby generating an acetyl hyaluronic acid oligopeptide.

[0011] The structure of acetylated hyaluronic acid is as shown in Formula I:

[0012] Formula I;

[0013] The structures of the oligopeptides are those of formulas II, III, and IV, respectively:

[0014] .

[0015] Preferably, it includes the following steps:

[0016] (1) Prepare acetyl hyaluronic acid-TBA from acetyl hyaluronic acid;

[0017] (2) Condensation reaction:

[0018] ① Solid-phase synthesis method: In the presence of an activated system and DIEA, CTC resin is used as the solid-phase synthesis carrier. Activated Fmoc-amino acids are sequentially condensed from the C-terminus to the N-terminus according to the amino acid sequences of oligopeptides of formulas II, III, and IV, respectively, to synthesize peptide resins. Three types of peptide resins with Fmoc-protecting groups at the N-terminus of oligopeptides of formulas II, III, and IV are synthesized. The sequence of Fmoc-amino acids sequentially condensed in the peptide resin of formula II of the synthesized oligopeptide is: Fmoc-Val-OH, Fmoc-Ala-OH, Fmoc-Val ... The synthetic oligopeptides are composed of the following structures: c-Lys(Dde)-OH, Fmoc-Ile-OH, and Fmoc-Ser-OH; the Fmoc-amino acids sequentially condensed by the III peptide resin are: Fmoc-Val-OH, Fmoc-Ala-OH, Fmoc-Val-OH, Fmoc-Lys(Dde)-OH, and Fmoc-Ile-OH; the synthetic oligopeptides are composed of the following structures: c-Lys(Dde)-OH, Fmoc-His-OH, and Fmoc-Gly-OH; the Fmoc-amino acids sequentially condensed by the IV peptide resin are: Fmoc-Lys(Dde)-OH, Fmoc-His-OH, and Fmoc-Gly-OH.

[0019] ② Remove the protecting group Dde of the Lys side chain ε-NH2 in the peptide resin containing the Fmoc protecting group obtained in step ① and cleave the peptide resin to obtain three kinds of crude oligopeptides with Fmoc protecting groups at the N-terminal amino groups of oligopeptide formula II, formula III and formula IV.

[0020] ③ The crude oligopeptide containing the Fmoc protecting group obtained in step ② was purified and lyophilized to obtain three pure oligopeptides with the N-terminal amino group containing the Fmoc protecting group of oligopeptide formula II, formula III and formula IV.

[0021] (3) After activating the acetyl hyaluronic acid-TBA obtained in step (1), it is condensed with the oligopeptide pure product obtained in step (2); the ε-NH2 of the Lys side chain in the oligopeptide sequence is connected with the -COOH of acetyl hyaluronic acid through an amide condensation reaction to generate an amide bond, and the product acetyl hyaluronic acid TBA oligopeptide-Fmoc is generated.

[0022] (4) Remove the Fmoc- protecting group of the N-terminal amino group of the oligopeptide in the product obtained in step (3) to obtain the product acetyl hyaluronic acid TBA oligopeptide;

[0023] (5) Salt exchange: The product acetyl hyaluronic acid TBA oligopeptide obtained in step (4) is ultrafiltered and washed through an ultrafiltration membrane (molecular weight cutoff 500 Da), and then converted to Na+ through a 732 type cation exchange column (resin). + Salt was freeze-dried to obtain acetyl hyaluronic acid oligopeptides with structures of formula V, formula VI and formula VII, respectively.

[0024] Preferably, the molecular weight of acetylated hyaluronic acid is 10 kDa to 100 kDa.

[0025] Preferably, the activation system in step (2) ① is A+D or A+B+C, where A is HOBT or HOAT, B is any one of HATU, HBTU, TBTU or PyBOP, C is DIEA or TMP, and D is DIC; the activation method of acetyl hyaluronic acid-TBA in step (3) is: acetyl hyaluronic acid-TBA reacts with HOSU in the presence of EDC for 24 h.

[0026] Preferably, the method for removing the protecting group Dde of the Lys side chain ε-NH2 in the peptide resin containing the Fmoc protecting group obtained in step (2) ② is as follows: add a deprotecting solution to the peptide resin containing the Fmoc protecting group obtained in step (2) ① and react for 3 hours. The deprotecting solution is hydroxylamine hydrochloride: imidazole: DCM: NMP = 25:18:16:100 (w / w / w / w).

[0027] Preferably, the method for lysing the peptide resin in step (2) ② is as follows: add 10 ml of lysis buffer per g of peptide resin, react at 10-30°C for 2-5 h to lyse it; the lysis buffer is 20% TFE / DCM (v / v).

[0028] Preferably, the purification method in step (2) ③ is to purify the crude oligopeptide containing the Fmoc protecting group obtained in step ② by reversed-phase high-performance liquid chromatography. The conditions are as follows: mobile phase A is H2O, mobile phase B is acetonitrile, gradient elution is used, the elution time is 60 min, the flow rate is 80 ml / min, the ultraviolet detection wavelength is 220 nm, and the elution gradient B phase is 10%~40% or 1%~31%.

[0029] The application method of any of the above-mentioned anti-wrinkle and anti-aging acetyl hyaluronic acid oligopeptides involves using any one, any two, or any three of the three acetyl hyaluronic acid oligopeptides having the structures of formula V, formula VI, and formula VII as a cosmetic component or adding them to a cosmetic.

[0030] The above-mentioned method of applying an anti-wrinkle and anti-aging acetyl hyaluronic acid oligopeptide involves using any one, two, or three of the three acetyl hyaluronic acid oligopeptides having structures of formula V, formula VI, and formula VII as a dermal filler in a facial injectable cosmetic product.

[0031] The beneficial effects of this invention are:

[0032] (1) The acetyl hyaluronic acid oligopeptide and its preparation and application method of the present invention, in view of the disadvantage of the unstable structure of acetyl hyaluronic acid and oligopeptide, the obtained acetyl hyaluronic acid oligopeptide has the functional activity of both acetyl hyaluronic acid and oligopeptide, has the targeting performance of acetyl hyaluronic acid to effectively promote the penetration of oligopeptide into the skin barrier, enhance the skin's absorption and binding of oligopeptide, increase the effect of oligopeptide use, and its moisturizing, wrinkle removal and anti-aging effects are significantly better than the effects of oligopeptide and acetyl hyaluronic acid used separately or in combination. At the same time, its chemical properties are stable, its anti-enzymatic ability is improved, its half-life is prolonged, it is not easily affected by the environment, its moisturizing, wrinkle removal and anti-aging effects are significant, and its action time is long.

[0033] (2) This invention uses covalent bonding to generate acetyl hyaluronic acid oligopeptide by coupling acetyl hyaluronic acid and oligopeptide, and directly chemically modifies it to give it better properties. The preparation and synthesis steps are simple and the purification is easy. At the same time, it will provide its application method in cosmetics and beauty products, significantly enhance the moisturizing, wrinkle removal and anti-aging effects of functional cosmetics, and significantly improve user satisfaction. Detailed Implementation

[0034] The present invention will be further described below with reference to specific embodiments to aid in understanding its content. Unless otherwise specified, the methods used in this invention are conventional production methods; and unless otherwise specified, the raw materials used are conventional commercially available products.

[0035] The specific meanings of the abbreviations used in this invention are listed in the table below:

[0036]

[0037] Example 1

[0038] Preparation of acetylated hyaluronic acid-TBA:

[0039] (1) Dissolve 300 g of acetylated hyaluronic acid with a molecular weight of 50 kDa in 10 L of deionized water, and then pass it through 732 (H + A type cation exchange column removes Na from the Na + Convert to H + The eluent containing acetylated hyaluronic acid was obtained.

[0040] (2) Add 200 g TBAOH to the eluent containing acetyl hyaluronic acid in step (1), stir for 2 h, and after neutralization, obtain acetyl hyaluronic acid-TBA reaction solution;

[0041] (3) The acetyl hyaluronic acid-TBA reaction solution obtained in step (2) was subjected to ultrafiltration, washing and concentration through an ultrafiltration membrane (molecular weight cutoff 500 D) to remove excess TBAOH and obtain acetyl hyaluronic acid-TBA product.

[0042] (4) The acetyl hyaluronic acid-TBA product obtained in step (3) is freeze-dried to obtain acetyl hyaluronic acid-TBA solid powder.

[0043] Example 2

[0044] Preparation of crude Fmoc-Ser-Ile-Lys-Val-Ala-Val-COOH peptide:

[0045] (1) Swelling of resin: Take 25 g of CTC resin with a substitution degree of 1.2 mmol / g, add 200 ml of DCM to swell the resin for 0.5 h, remove the solvent, wash the resin twice with DMF, and remove the solvent.

[0046] (2) Preparation of Fmoc-Val-CTC resin: a) Fmoc-Val-OH, DIEA and CTC resin swollen in step (1) were mixed in a molar mass ratio of 3:6:1 and reacted at 25°C for 2 h to obtain Fmoc-Val-CTC resin; b) A mixed solution of MeOH, DMF and DIEA was added to the resin and reacted at 30°C for 30 min to block the resin. The resin was then washed twice with DMF and the solvent was removed to obtain the blocked Fmoc-Val-CTC resin.

[0047] (3) Removal of Fmoc protecting groups: Add 20% PIP-DMF solution (volume fraction) to the blocked Fmoc-Val-CTC resin obtained in step (2), and perform two Fmoc protection removals at 10-30℃: After the first and second Fmoc protection removals, wash the resin with DMF until the pH is 7; the first Fmoc protection removal time is 5 min, and the second Fmoc protection removal time is 10 min, to obtain resin with Fmoc protecting groups removed;

[0048] (4) Amino acid activation: At room temperature, 90 mmol of each of Fmoc-Ala-OH, Fmoc-Val-OH, Fmoc-Lys(Dde)-OH, Fmoc-Ile-OH and Fmoc-Ser-OH and 90 mmol of HOBT were dissolved in an appropriate amount of DMF, and then 90 mmol of DIC was added. The activation reaction was carried out for 5 min to obtain activated amino acids.

[0049] (5) Amino acid condensation: The activated Fmoc-Ala-OH, Fmoc-Val-OH, Fmoc-Lys(Dde)-OH, Fmoc-Ile-OH and Fmoc-Ser-OH obtained in step (4) are added to the resin obtained in step (3) after the Fmoc protecting group is removed. The amino acid condensation reaction is carried out at 25°C for 2 h. The reaction process is monitored by ninhydrin colorimetric reaction to obtain Fmoc-Ser-Ile-Lys(Dde)-Val-Ala-Val-CTC resin.

[0050] (6) Removal of the Dde protecting group: The Fmoc-Ser-Ile-Lys(Dde)- obtained in step (5) is deprotected.

[0051] In Val-Ala-Val-CTC resin, add the deprotecting solution hydroxylamine hydrochloride:imidazole:DCM:NMP = 25:18:

[0052] After reacting at a ratio of 16:100 (w / w / w / w) for 3 h, the peptide resin was washed three times with 300 ml DMF and 300 ml MeOH alternately for 5 min each time. The peptide resin was then shrunken, the solvent was removed, and the resin was dried under vacuum to obtain Fmoc-Ser-Ile-Lys-Val-Ala-Val-CTC resin.

[0053] (7) Lysis: Add 10 ml of lysis buffer per gram of peptide resin to a round-bottom flask.

[0054] The ratio of TFE / DCM (v / v) to the Fmoc-Ser-Ile-Lys-Val-Ala-Val obtained in step (6)

[0055] CTC resin was pyrolyzed and reacted at 25°C for 2 h. The mixture was then filtered, and the filtrate was subjected to vacuum distillation to evaporate the solvent, yielding crude Fmoc-Ser-Ile-Lys-Val-Ala-Val-COOH peptide.

[0056] Example 3

[0057] Preparation of crude Fmoc-Ile-Lys-Val-Ala-Val-COOH peptide:

[0058] (1) Swelling of resin: Take 25 g of CTC resin with a substitution degree of 1.2 mmol / g, add 200 ml of DCM to swell the resin for 0.5 h, remove the solvent, wash the resin twice with DMF, and remove the solvent.

[0059] (2) Preparation of Fmoc-Val-CTC resin: a) Fmoc-Val-OH, DIEA and resin were mixed in a molar mass ratio of 3:6:1 and reacted at 25℃ for 2 h to obtain Fmoc-Val-CTC resin; b) A mixed solution of MeOH, DMF and DIEA was added to the resin and reacted at 10-30℃ for 30 min to block the resin. The resin was then washed twice with DMF and the solvent was removed to obtain the blocked Fmoc-Val-CTC resin.

[0060] (3) Removal of Fmoc protecting groups: Add 20% PIP-DMF solution (volume fraction) to the blocked Fmoc-Val-CTC resin obtained in step (2), and perform two Fmoc protection removals at 10~30℃: After the first and second Fmoc protection removals, wash the resin with DMF until the pH is 7; the first Fmoc protection removal time is 5 min, and the second Fmoc protection removal time is 10 min, to obtain the resin with Fmoc protecting groups removed;

[0061] (4) Amino acid activation: 90 mmol of each of Fmoc-Ala-OH, Fmoc-Val-OH, Fmoc-Lys(Dde)-OH and Fmoc-Ile-OH and 90 mmol of HOBT were dissolved in an appropriate amount of DMF, and then 90 mmol of DIC was added. The reaction was carried out at room temperature for 5 min to obtain activated amino acids.

[0062] (5) Amino acid condensation: The activated Fmoc-Ala-OH, Fmoc-Val-OH, Fmoc-Lys(Dde)-OH and Fmoc-Ile-OH from step (4) were added sequentially to the resin after the Fmoc protecting group was removed, and the amino acid condensation reaction was carried out at 25°C for 2 h. The reaction process was monitored by the ninhydrin color reaction, and finally Fmoc-Ile-Lys(Dde)-Val-Ala-Val-CTC resin was obtained.

[0063] (6) Removal of the Dde protecting group: The Fmoc-Ile-Lys(Dde)- obtained in step (5) is deprotected.

[0064] Add the deprotection solution hydroxylamine hydrochloride:imidazolium:DCM:NMP = 25:18:16:100 (w / w / w / w) to Val-Ala-Val-CTC resin. After reacting for 3 h, wash the peptide resin three times alternately with 300 ml DMF and 300 ml MeOH for 5 min each time. Shrink the peptide resin, remove the solvent, and vacuum dry to obtain Fmoc-Ile-Lys-Val-Ala-Val-CTC resin.

[0065] (7) Cleavage: The Fmoc-Ile-Lys-Val-Ala-Val-CTC resin obtained in step (6) was cleaved in a round-bottom flask at a ratio of 10 ml of 20% TFE / DCM (v / v) per g of peptide resin. The reaction was carried out at 25°C for 2 h. The mixture was then filtered, and the filtrate was subjected to vacuum distillation to evaporate the solvent, thereby obtaining crude Fmoc-Ile-Lys-Val-Ala-Val-COOH peptide.

[0066] Example 4

[0067] Preparation of crude Fmoc-Gly-His-Lys-COOH peptide:

[0068] (1) Swelling of resin: Take 37.5 g of CTC resin with a substitution degree of 1.2 mmol / g, add 300 ml of DCM to swell the resin for 0.5 h, remove the solvent, wash the resin twice with DMF, and remove the solvent.

[0069] (2) Preparation of Fmoc-Lys(Dde)-CTC resin: a) Fmoc-Lys(Dde)-OH, DIEA and resin were mixed in a molar mass ratio of 3:6:1 and reacted at 25℃ for 2h to obtain Fmoc-Lys(Dde)-CTC resin; b) A mixed solution of MeOH, DMF and DIEA was added to the resin and reacted at 10~30℃ for 30 min to block the resin. The resin was then washed twice with DMF and the solvent was removed to obtain the blocked Fmoc-Lys(Dde)-CTC resin.

[0070] (3) Removal of Fmoc protecting groups: Add 20% PIP-DMF solution (by volume) to the blocked Fmoc-Lys(Dde)-CTC resin obtained in step (2), and perform two Fmoc protection removals at 10-30°C: After the first and second Fmoc protection removals, wash the resin with DMF until the pH is 7; the first Fmoc protection removal time is 5 min, and the second Fmoc protection removal time is 10 min, to obtain resin with Fmoc protecting groups removed;

[0071] (4) Amino acid activation: 135 mmol each of Fmoc-His-OH and Fmoc-Gly-OH and 135 mmol of HOBT were dissolved in an appropriate amount of DMF, and then 135 mmol of DIC was added. The reaction was carried out at room temperature for 5 min to obtain activated amino acids.

[0072] (5) Amino acid condensation: The activated Fmoc-His-OH and Fmoc-Gly-OH from step (4) were added sequentially to the resin after the Fmoc protecting group was removed, and the amino acid condensation reaction was carried out at 25°C for 2 hours. The reaction process was monitored by the ninhydrin color reaction, and finally Fmoc-Gly-His-Lys-CTC resin was obtained.

[0073] (6) Removal of Dde protecting group: Add 300 ml of deprotection solution hydroxylamine hydrochloride: imidazole: DCM: NMP = 25:18:16:100 (w / w / w / w) to the Fmoc-Gly-His-Lys-CTC resin obtained in step (5). After reacting for 3 h, wash the peptide resin three times with 300 ml DMF and 300 ml MeOH alternately for 5 min each time. Shrink the peptide resin, remove the solvent, and vacuum dry to obtain Fmoc-Gly-His-Lys-CTC resin.

[0074] (7) Lysis: Add 10 ml of lysis buffer per gram of peptide resin to a round-bottom flask.

[0075] The ratio of TFE / DCM (v / v) was used to cleave the Fmoc-Gly-His-Lys-CTC resin obtained in step (6). The reaction was carried out at 25°C for 2 hours. The mixture was then filtered, and the filtrate was subjected to vacuum distillation to evaporate the solvent, thereby obtaining crude Fmoc-Gly-His-Lys-COOH peptide.

[0076] Example 5

[0077] purification:

[0078] The crude peptides of Fmoc-Ser-Ile-Lys-Val-Ala-Val-COOH, Fmoc-Ile-Lys-Val-Ala-Val-COOH, and Fmoc-Gly-His-Lys-COOH prepared in Examples 2-4 were purified respectively.

[0079] Three crude peptides were purified in one step using reversed-phase high-performance liquid chromatography (RP-HPLC). Mobile phase A was H₂O, and mobile phase B was acetonitrile. Gradient elution was employed: a C18 preparative column (50 × 250 mm, 10 µm) was used, with an elution time of 60 min. The elution gradient for the oligopeptides obtained in Examples 2 and 3 was 10%–40% in phase B; the elution gradient for the oligopeptides obtained in Example 4 was 1%–31% in phase B. The flow rate was 80 ml / min, and the UV detection wavelength was 220 nm. Finally, the peptides were concentrated and lyophilized to obtain Fmoc-Ser-Ile-Lys-Val-Ala-Val-COOH peptide, Fmoc-Ile-Lys-Val-Ala-Val-

[0080] The purities of the three oligopeptides, COOH peptide, Fmoc-Gly-His-Lys-COOH peptide, were 98.2%, 98.0%, and 98.3%, respectively.

[0081] Example 6

[0082] Preparation of acetylhyaluronic acid oligopeptide product:

[0083] (1) Activation of acetylated hyaluronic acid: 50 g of acetylated hyaluronic acid prepared in Example 1 was activated.

[0084] TBA was dissolved in DMF, and 17 mmol HOSU and 34 mmol EDC were added. The mixture was reacted at 30°C for 24 h. The reaction solution was poured into 4 times the volume of ethyl acetate, stirred for 0.5 h, and then allowed to stand overnight to allow precipitation. The precipitate was filtered and washed twice with ethyl acetate to obtain activated acetyl hyaluronic acid.

[0085] (2) Dissolve the activated acetyl hyaluronic acid obtained in step (1) in DMF, add 17 mmol of any one of the three oligopeptide pure products prepared in Example 5: Fmoc-Ser-Ile-Lys-Val-Ala-Val-COOH peptide, Fmoc-Ile-Lys-Val-Ala-Val-COOH peptide, and Fmoc-Gly-His-Lys-COOH peptide, and add 20.4 mmol of DIEA to stir the reaction. Monitor the reaction progress with ninhydrin colorimetric reaction until there are no amino components in the reaction solution to make the reaction complete, and obtain Fmoc protected acetyl hyaluronic acid oligopeptide;

[0086] (3) Add 10% by volume of diethylamine to the reaction system of Fmoc-protected acetyl hyaluronic acid oligopeptide obtained in step (2) to remove the Fmoc protecting group at the N end of the peptide chain, and then evaporate the solvent by vacuum distillation to obtain the semi-finished product of acetyl hyaluronic acid oligopeptide.

[0087] (4) Dissolve the acetyl hyaluronic acid oligopeptide semi-finished product obtained in step (3), perform ultrafiltration and washing through an ultrafiltration membrane (molecular weight cutoff 500D), and then convert the TBA in it to Na through a 732 cation exchange column. + Three types of acetyl hyaluronic acid oligopeptides with structures of formula V, formula VI and formula VII were obtained by freeze drying.

[0088] Example 7

[0089] Preparation of acetylhyaluronic acid oligopeptide product:

[0090] (1) 50 g of acetylated hyaluronic acid-TBA prepared in Example 1 was dissolved in DMF, 34 mmol HOSU and 68 mmol EDC were added, and the mixture was reacted at 30°C for 24 h. The reaction solution was poured into 4 times the volume of ethyl acetate, stirred for 0.5 h, and allowed to stand overnight for precipitation. The precipitate was filtered and washed twice with ethyl acetate to obtain activated acetylated hyaluronic acid.

[0091] (2) Dissolve the activated acetylated hyaluronic acid obtained in step 5 in DMF, add 34 mmol of any one of the three oligopeptide pure products prepared in Example 5: Fmoc-Ser-Ile-Lys-Val-Ala-Val-COOH peptide, Fmoc-Ile-Lys-Val-Ala-Val-COOH peptide, and Fmoc-Gly-His-Lys-COOH peptide, and add 40.8 mmol of DIEA to stir the reaction. Monitor the reaction process with ninhydrin colorimetric reaction until there are no amino components in the reaction solution to make the reaction complete, and obtain Fmoc protected acetylated hyaluronic acid oligopeptide;

[0092] (3) Add 10% by volume of diethylamine to the reaction system of Fmoc-protected acetyl hyaluronic acid oligopeptide obtained in step (2) to remove the Fmoc protecting group at the N end of the peptide chain, and then evaporate the solvent by vacuum distillation to obtain the semi-finished product of acetyl hyaluronic acid oligopeptide.

[0093] (4) Dissolve the acetyl hyaluronic acid oligopeptide semi-finished product obtained in step (3), perform ultrafiltration and washing through an ultrafiltration membrane (molecular weight cutoff 500D), and then convert the TBA in it to Na through a 732 cation exchange column. + Three types of acetyl hyaluronic acid oligopeptides with structures of formula V, formula VI and formula VII were obtained by freeze drying.

[0094] Example 8

[0095] Preparation of acetylhyaluronic acid oligopeptide product:

[0096] (1) Dissolve 50 g of acetylated hyaluronic acid-TBA prepared in Example 1 in DMF, add 51 mmol HOSU and 102 mmol EDC, react at 30°C for 24 h, pour the reaction solution into 4 times the volume of ethyl acetate, stir for 0.5 h and let stand overnight to allow precipitation; filter the precipitate and wash it twice with ethyl acetate to obtain activated acetylated hyaluronic acid;

[0097] (2) Dissolve the activated acetyl hyaluronic acid obtained in step (1) in DMF, add 51 mmol of any one of the three oligopeptide pure products prepared in Example 5: Fmoc-Ser-Ile-Lys-Val-Ala-Val-COOH peptide, Fmoc-Ile-Lys-Val-Ala-Val-COOH peptide, and Fmoc-Gly-His-Lys-COOH peptide, and add 61.2 mmol of DIEA to stir the reaction. Use ninhydrin colorimetric reaction to monitor the reaction process until there are no amino components in the reaction solution to make the reaction complete, and obtain Fmoc protected acetyl hyaluronic acid oligopeptide;

[0098] (3) Add 10% by volume of diethylamine to the reaction system of Fmoc-protected acetyl hyaluronic acid oligopeptide obtained in step (2) to remove the Fmoc protecting group at the N end of the peptide chain, and then evaporate the solvent by vacuum distillation to obtain the semi-finished product of acetyl hyaluronic acid oligopeptide.

[0099] (4) Dissolve the solid obtained in step (3), perform ultrafiltration and washing through an ultrafiltration membrane (molecular weight cutoff 500D), and then convert the TBA in it to Na through a 732 cation exchange column. + Three types of acetyl hyaluronic acid oligopeptides with structures of formula V, formula VI and formula VII were obtained by freeze drying.

[0100] Example 9

[0101] A moisturizing and anti-wrinkle cosmetic serum containing acetylated hyaluronic acid oligopeptide as a component:

[0102] At room temperature, 4% butylene glycol and 0.8% PE9010 were added to seven equal portions of sterile water as matrices, respectively, serving as experimental groups 1-6 and control group 7. Then, 0.5% acetylated hyaluronic acid, oligopeptide, a combination of acetylated hyaluronic acid and oligopeptide, and three acetylated hyaluronic acid oligopeptide products with structures of formulas V, VI, and VII prepared in Example 8 were added to experimental groups 1-6, respectively. An equal mass of sterile water was added to control group 7, and all mixtures were stirred until complete. After complete dissolution and adjustment of pH to 5-7, experimental groups 1 (hyaluronic acid stock solution), 2 (oligopeptide stock solution containing three types of oligopeptides with structures of formula II, III, and IV), 3 (acetyl hyaluronic acid combined with oligopeptides (three types of oligopeptides with structures of formula II, III, and IV), 4 (acetyl hyaluronic acid oligopeptide stock solution with structure V), 5 (acetyl hyaluronic acid oligopeptide stock solution with structure VI), 6 (acetyl hyaluronic acid oligopeptide stock solution with structure VII), and 7 (blank group) were obtained.

[0103] The performance of the acetyl hyaluronic acid oligopeptide of the present invention will be further illustrated below through experimental reports.

[0104] (1) Testing items:

[0105] The moisturizing and wrinkle-reducing effects of acetylated hyaluronic acid oligopeptide were tested.

[0106] (2) Test sample:

[0107] The moisturizing and wrinkle-reducing effects of the hyaluronic acid stock solution prepared in Example 9, experimental group 1, experimental group 2 (three oligopeptides with structures of formula II, formula III and formula IV respectively), experimental group 3 (acetyl hyaluronic acid and oligopeptides (three oligopeptides with structures of formula II, formula III and formula IV respectively) combination stock solution, experimental group 4 (acetyl hyaluronic acid oligopeptide stock solution with structure V), experimental group 5 (acetyl hyaluronic acid oligopeptide stock solution with structure VI), experimental group 6 (acetyl hyaluronic acid oligopeptide stock solution with structure VII), and blank group 7 are shown in Tables 1-3.

[0108] (3) Determination of the efficacy of acetyl hyaluronic acid oligopeptide:

[0109] 1. Moisturizing effect:

[0110] 1.1 Subjects: 110 volunteers aged 35-40 years were selected and their forearm skin was tested.

[0111] 1.2 Test Samples: The original solutions of experimental groups 1-6 and blank group 7 prepared in Example 9 were used as test samples;

[0112] 1.3 Before the experiment: Subjects were required to wash the inside of their forearms with clean water. After washing, measurement marks were made on the inside of both forearms. In this experiment, two test areas were marked on each forearm.

[0113] 1.4 In the experiment: Seven test areas were divided on each test subject's arm, with a 1cm interval between areas, and each test area was 3×3cm. 2 The sample size was 0.2g. Each subject applied one test substance to each area, and the stock solutions of experimental groups 1-6 and blank group 7 prepared in Example 9 were applied to seven test areas. Subjects sat quietly for 30 minutes in a constant environment (measurement temperature 20℃, relative humidity 50%). Blank values ​​were measured at the test sites using a Corneometer CM 825 moisture analyzer. Five points were measured in a fixed order for each area, and the average value was obtained. Then, a designated person applied the sample and started timing, measuring the changes in MMV values ​​at various time points according to the experimental design. The change in MMV value for each test area was obtained by subtracting the blank value from the average value measured each time; further subtracting the blank value yielded the growth rate of the MMV value.

[0114] Average moisture content growth rate % = (MMV) t -MMV0) / MMV0×100%,

[0115] In the formula: MMV0—the skin's MMV before application.

[0116] MMV t —Skin MMV at time t after application.

[0117] Table 1 shows the test experience reference data for this testing method: The following data were obtained under normal room temperature conditions (measurement ambient temperature is 20℃, relative humidity is 50%).

[0118] Table 1. Test experience data of Corneometer CM 825 moisture analyzer

[0119]

[0120] The moisture content of experimental groups 1-6 and blank group 7 prepared in Example 9 was tested using a Corneometer CM 825 moisture analyzer within 4 hours. The average value of the change in skin MMV value of each test substance was calculated by testing the skin of the forearms of 70 volunteers. The data results are shown in Table 2.

[0121]

[0122] The experimental groups included: hyaluronic acid stock solution (Group 1), oligopeptide stock solution (three types of oligopeptides with structures of formulas II, III, and IV) (Group 2), acetyl hyaluronic acid and oligopeptide combination stock solution (Group 3), acetyl hyaluronic acid oligopeptide stock solution with structure V (Group 4), acetyl hyaluronic acid oligopeptide stock solution with structure VI (Group 5), acetyl hyaluronic acid oligopeptide stock solution with structure VII (Group 6), and a blank group (Group 7). The moisturizing effect test results for the three types of oligopeptides with structures II, III, and IV in Groups 2 and 3 were similar. Therefore, the moisturizing effect test data for the oligopeptide stock solution in Group 2 used in Table 1 is the average of the moisturizing effect test data for the three types of oligopeptides. The moisturizing effect test data for the acetyl hyaluronic acid and oligopeptide combination stock solution in Group 3 is also the average of the moisturizing effect test data for the mixture of the three types of oligopeptides and acetyl hyaluronic acid.

[0123] Table 1, comparing the moisturizing effects of acetyl hyaluronic acid oligopeptides, shows that the skin's MMV value generally decreases over time. The blank group 7 exhibited the worst moisturizing effect, demonstrating that without added hydrating ingredients, the skin cannot effectively retain moisture when only a sterile water matrix is ​​provided. The moisturizing effects of experimental groups 4-6 (acetyl hyaluronic acid oligopeptide stock solution) were significantly higher than those of experimental group 1 (hyaluronic acid stock solution) and experimental group 2 (oligopeptide stock solution), and also significantly higher than those of experimental group 3 (acetyl hyaluronic acid and oligopeptide combination stock solution). This demonstrates that the significant improvement in the moisturizing effect of the acetyl hyaluronic acid oligopeptide prepared in this invention is not due to a simple combination of acetyl hyaluronic acid and oligopeptide, but rather because acetyl hyaluronic acid and oligopeptide are covalently coupled to form acetyl hyaluronic acid oligopeptide. This chemical modification fundamentally endows it with superior properties, improving the stability of acetyl hyaluronic acid and oligopeptide, enhancing their resistance to enzymatic degradation, prolonging their half-life, and increasing their duration of action, thus significantly improving their moisturizing effect. In particular, experimental groups 4-6 showed an upward trend in skin MMV values ​​at 120 minutes, which fully demonstrates that the effect of acetylated hyaluronic acid oligopeptide is not limited to the skin surface, but also has a significant effect on the hydration and repair of deep cells, ultimately resulting in a slow decrease in skin moisture and achieving a long-lasting moisturizing effect.

[0124] 2. Wrinkle-reducing effect:

[0125] 2.1 Subjects: Seventy volunteers aged 35-55 years were selected and divided into 7 groups of 10 people each to conduct skin experiments;

[0126] 2.2 Test samples and their usage: 0.5g of each of the stock solutions of experimental groups 1-6 and blank group 7 prepared in Example 9 were evenly applied to the facial skin of volunteers once a day for 4 consecutive weeks.

[0127] Before the experiment, the area of ​​facial skin wrinkles S0 of the subjects was measured using the Visioline VL 650 wrinkle tester. The changes in the area of ​​facial skin wrinkles were obtained by analyzing the changes using software.

[0128] In the experiment, at 2 PM each day, with an ambient temperature of 20℃ and relative humidity of 50%, the area of ​​facial wrinkles (S) was measured using a Visioline VL 650 wrinkle analyzer. t Changes in the silicone replica membrane of skin wrinkles were analyzed using software to determine the changes in the area of ​​facial skin wrinkles.

[0129] Finally, calculate the reduction in skin wrinkle area (%) = (pre-experiment skin wrinkle area - average weekly skin wrinkle area) / pre-experiment skin wrinkle area × 100% = (S0 - S t ) / S0×100%.

[0130]

[0131] The experimental groups included: hyaluronic acid stock solution (Group 1), oligopeptide stock solution (Group 2, consisting of three oligopeptides with structures of formulas II, III, and IV) (Group 2), acetyl hyaluronic acid and oligopeptides (Group 3, consisting of three oligopeptides with structures of formulas II, III, and IV) (Group 3), acetyl hyaluronic acid oligopeptide stock solution with structure V (Group 4), acetyl hyaluronic acid oligopeptide stock solution with structure VI (Group 5), acetyl hyaluronic acid oligopeptide stock solution with structure VII (Group 6), and a blank group (Group 7). The wrinkle-reducing effect data of the three oligopeptides with structures II, III, and IV in Groups 2 and 3 were similar. Therefore, the wrinkle-reducing effect data of the oligopeptide stock solution in Group 2 used in Table 1 are the average values ​​of the wrinkle-reducing effect data of the three oligopeptide structures. The wrinkle-reducing effect data of the acetyl hyaluronic acid and oligopeptide combination stock solution in Group 3 are also the average values ​​of the wrinkle-reducing effect data of the mixture of the three oligopeptides with acetyl hyaluronic acid.

[0132] Table 2, comparing the wrinkle-reducing effects of acetyl hyaluronic acid oligopeptides, shows that the reduction in wrinkle area in the blank group 7 remained almost unchanged, proving that wrinkles on the human face do not disappear on their own once formed without the addition of anti-wrinkle products. The wrinkle-reducing effects of the acetyl hyaluronic acid oligopeptide stock solution in experimental groups 4-6 were significantly higher than those of the hyaluronic acid stock solution in experimental group 1 and the oligopeptide stock solution in experimental group 2 alone, and also significantly higher than the combination stock solution of acetyl hyaluronic acid and oligopeptides in experimental group 3. This fully demonstrates the significant improvement in wrinkle-reducing effect of the acetyl hyaluronic acid oligopeptide prepared in this invention. The high concentration of acetylated hyaluronic acid is not simply a result of the combination of acetylated hyaluronic acid and oligopeptides. Rather, it is due to the covalent coupling of acetylated hyaluronic acid and oligopeptides to form acetylated hyaluronic acid oligopeptides. This direct chemical modification fundamentally endows them with superior properties, improving the stability of both acetylated hyaluronic acid and oligopeptides, enhancing their resistance to enzymatic degradation, prolonging their half-life, and extending their duration of action. Acetylated hyaluronic acid oligopeptides, possessing the targeting properties of acetylated hyaluronic acid, effectively promote the penetration of oligopeptides into the skin barrier, enhancing the skin's absorption and binding of oligopeptides, thus increasing the effectiveness of oligopeptides and significantly improving their wrinkle-reducing effect. Preferably, the moisturizing and wrinkle-reducing effect is most significant when the added amount of acetylated hyaluronic acid oligopeptides is between 0.001% and 25%.

[0133] This method synthesizes oligopeptides and acetylated hyaluronic acid by covalently linking them through chemical modification. The resulting acetylated hyaluronic acid oligopeptide exhibits complex activity, strong efficacy, and long-lasting effects. Its efficacy surpasses that of either acetylated hyaluronic acid or oligopeptide alone, and also outperforms the combined use of acetylated hyaluronic acid and oligopeptide. The method for its application in cosmetics and beauty products is also provided.

[0134] In summary, this invention addresses the structural instability of acetylated hyaluronic acid and oligopeptides, solving the technical problems of existing functional cosmetics where acetylated hyaluronic acid and oligopeptides need to be added separately, are easily hydrolyzed by enzymes, have poor chemical stability, are easily inactivated by environmental influences, and have short durations of action. The prepared acetylated hyaluronic acid oligopeptide combines the functional activities of both acetylated hyaluronic acid and oligopeptides, and its wrinkle-reducing and anti-aging effects are significantly better than those of oligopeptides and acetylated hyaluronic acid used separately or in combination. It also prolongs the efficacy, enhances the effectiveness of functional cosmetics, improves user satisfaction, and has a wide range of applications.

[0135] The amount of acetyl hyaluronic acid oligopeptide added; the application scope of acetyl hyaluronic acid oligopeptide: it can also be used to formulate cosmetics with functions such as sun protection, moisturizing, skin nutrition replenishment, anti-inflammation, anti-oxidation, wrinkle removal, anti-aging and skin repair; the dosage forms of cosmetics include aqueous solutions, lotions, essences, gels, foundations, creams and masks; the application scope of cosmetics includes head care, facial care and body care, etc., all of which can realize the application method of acetyl hyaluronic acid oligopeptide of the present invention.

[0136] The above are merely embodiments of the present invention. For example, the molecular weight of acetyl hyaluronic acid is 10 kDa to 100 kDa; the activation system in step (2) ① of the condensation reaction is A+D or A+B+C, where A is HOBT or HOAT, B is any one of HATU, HBTU, TBTU or PyBOP, C is DIEA or TMP, and D is DIC; the method of cleaving peptide resin is to add 10 ml of cleavage solution per g of peptide resin, react at 10 to 30°C for 2-5 h, and cleave it, etc., to realize the acetyl hyaluronic acid oligopeptide of the present invention and its preparation.

[0137] However, the above description is merely a specific embodiment of the present invention and should not be construed as limiting the scope of the present invention. All equivalent changes and modifications made in accordance with the claims of the present invention should be covered by the present invention.

Claims

1. A hyaluronan oligopeptide, characterized in that, The structure of the hyaluronic acid oligopeptide is a glycopeptide with the structure of formula VII: 。 2. The method for preparing hyaluronic acid oligopeptide according to claim 1, characterized in that, The method comprises the following steps: The hyaluronic acid is connected with the oligopeptide in the form of a covalent bond: the ε-NH2 of the Lys side chain in the oligopeptide sequence is connected with the -COOH of the hyaluronic acid through an amide condensation reaction to form an amide bond, thereby generating the hyaluronic acid oligopeptide; The structure of the hyaluronic acid is formula I: Formula I; The structure of the oligopeptide is formula IV: 。 3. The method for preparing hyaluronic acid oligopeptide according to claim 2, characterized in that, The method comprises the following steps: (1) The hyaluronic acid is prepared into hyaluronic acid-TBA; (2) Condensation reaction: ① By using a solid-phase synthesis method: in the presence of an activation system and DIEA, the CTC resin is used as a solid-phase synthesis carrier, and the activated Fmoc-amino acid is sequentially condensed according to the amino acid sequence of the oligopeptide of formula IV from the C-terminal to the N-terminal, thereby synthesizing the peptide resin of the N-terminal amino group of the oligopeptide of formula IV containing a Fmoc protection group; the sequentially condensed Fmoc-amino acid sequence of the peptide resin of the synthesized oligopeptide of formula IV is: Fmoc-Lys(Dde)-OH, Fmoc-His-OH, Fmoc-Gly-OH; ② The protection group Dde of the Lys side chain ε-NH2 in the Fmoc-protected peptide resin prepared in step ① is removed, and the peptide resin is cleaved, thereby obtaining the crude oligopeptide of formula IV whose N-terminal amino group contains a Fmoc protection group; ③ The Fmoc-protected crude oligopeptide prepared in step ② is purified and freeze-dried, thereby obtaining the Fmoc-protected oligopeptide product of formula IV whose N-terminal amino group contains a Fmoc protection group; (3) The hyaluronic acid-TBA prepared in step (1) is activated and subjected to a condensation reaction with the oligopeptide product prepared in step (2); the ε-NH2 of the Lys side chain in the oligopeptide sequence is connected with the -COOH of the hyaluronic acid through an amide condensation reaction to form an amide bond, thereby generating the product hyaluronic acid TBA oligopeptide-Fmoc; (4) The Fmoc protection group of the N-terminal amino group of the product prepared in step (3) is removed, thereby obtaining the product hyaluronic acid TBA oligopeptide; (5) Salt exchange: the product hyaluronic acid TBA oligopeptide prepared in step (4) is subjected to ultrafiltration and washing through an ultrafiltration membrane with a molecular weight cut-off of 500 Da, and then converted into a Na + salt through a 732 type cation exchange column resin, and then lyophilized to obtain a hyaluronic acid oligopeptide finished product with the structure of formula VII.

4. The method for preparing hyaluronic acid oligopeptide according to claim 3, characterized in that, The molecular weight of the hyaluronic acid is 10 kDa to 100 kDa.

5. The method for preparing an acetylhyaluronic acid oligopeptide according to claim 3, characterized in that, In step (2) ①, the activation system is A+D or A+B+C, wherein A is HOBT or HOAT, B is any one of HATU, HBTU, TBTU or PyBOP, C is DIEA or TMP, and D is DIC; in step (3), the activation method of the hyaluronic acid-TBA is that the hyaluronic acid-TBA is reacted with HOSU in the presence of EDC for 24 h.

6. The method for preparing an acetylhyaluronic acid oligopeptide according to claim 3, characterized in that, In step (2) ②, the removal method of the protection group Dde of the Lys side chain ε-NH2 in the Fmoc-protected peptide resin prepared in step ① is that the Fmoc-protected peptide resin prepared in step (2) ① is added with a deprotection solution for 3 h, and the deprotection solution is composed of hydroxylamine hydrochloride, imidazole, DCM and NMP, and the mass ratio of hydroxylamine hydrochloride, imidazole, DCM and NMP is 25:18:16:

100.

7. The method for preparing an acetylhyaluronic acid oligopeptide according to claim 3, characterized in that, The method for cleaving the peptide resin in step (2) ii) is as follows: 10 ml of cleavage solution is added per gram of peptide resin, and the reaction is carried out at 10-30°C for 2-5 h to cleave the peptide resin; the cleavage solution is composed of TFE and DCM, and the volume ratio of TFE to DCM is 2:

8.

8. The method for preparing an acetylhyaluronic acid oligopeptide according to claim 3, characterized in that, The purification method in step (2) iii) is as follows: the Fmoc-protected oligopeptide crude peptide prepared in step (2) ii) is purified by reverse-phase high-performance liquid chromatography, and the conditions are as follows: mobile phase A is H2O, mobile phase B is acetonitrile, gradient elution is used, the elution time is 60 min, the flow rate is 80 ml / min, and the ultraviolet detection wavelength is 220 nm; the elution gradient B is 1%-31%.

9. Use of a hyaluronic acid oligopeptide according to claim 1 or produced by the method according to any one of claims 2 to 8, characterized in that, The hyaluronic acid oligopeptide having the structure of formula VII is used as a cosmetic component or is added to a cosmetic product.

10. Use of a hyaluronic acid oligopeptide according to claim 1 or produced by the method according to any one of claims 2 to 8, characterized in that, The hyaluronic acid oligopeptide having the structure of formula VII is used as a skin filler to prepare a facial injection plastic and cosmetic product.

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