Application of oligopeptides in improving the stability of cross-linked hyaluronic acid or its salts

Abstract

The present application provides an application of an oligopeptide in improving the stability of cross-linked hyaluronic acid or its salt, and an injectable hyaluronic acid gel. The viscosity of cross-linked hyaluronic acid decreases after sterilization, and the addition of oligopeptides at a certain mass ratio can alleviate the decrease in viscosity of cross-linked hyaluronic acid and maintain the stability of cross-linked hyaluronic acid after sterilization. Using cross-linked hyaluronic acid gel as a matrix and compounding other nutrients can replenish water and moisturize, stimulate fibroblast activity, increase fibroblast synthesis of substrates, and thus improve the effect of skin quality.

Description

Technical Field

[0001] The present application relates to an oligopeptide, and specifically to an application of an oligopeptide in improving the stability of cross-linked hyaluronic acid or its salt, and an injectable hyaluronic acid gel. Background Art

[0002] With economic development and social progress, people pay more and more attention to their appearance. The skin is the outermost and largest organ of the human body. Like other organs of the human body, it will gradually age, weaken or lose its functions, which not only affects the appearance, but also increases the incidence of various skin diseases. In addition, due to its superficial exposure, the skin has a strong psychological effect, and delaying skin aging has gradually become one of the hot topics of people's attention. In particular, minimally invasive facial beauty rejuvenation is sought after by many beauty seekers and is also one of the research hotspots in the field of life sciences at home and abroad.

[0003] Water injection is a commonly used minimally invasive cosmetic method that can directly inject nutrients needed by the skin into the skin. After being directly and quickly absorbed by the subcutaneous tissue, it has the effect of anti-aging and regeneration of the skin. Hyaluronic acid (HA) is a commonly used water injection material.

[0004] Hyaluronic acid is a linear high molecular mucopolysaccharide composed of repeated disaccharide units of glucuronic acid and acetylglucosamine. Hyaluronic acid is an endogenous substance in the human body and has good biocompatibility. As an important component of the extracellular matrix, hyaluronic acid has superb hydration, repair, anti-aging and wrinkle removal capabilities. Hyaluronic acid can lock 1000 times its own weight of water in the body and is the best moisturizing substance found in nature so far. The main cause of skin aging is lack of water, and the root cause of lack of water is the loss of hyaluronic acid. Therefore, replenishing hyaluronic acid through the skin to effectively replenish skin moisture is a fundamental solution to solve skin aging. The water-light injection product with hyaluronic acid gel as the matrix and compounded with other nutrients can not only play a hydrating and moisturizing effect, but also can locate, determine the layers, and quantitatively transport skin nutrients containing multiple nutrients and high concentrations directly through the physiological barrier of the epidermis to the dermis and deep subcutaneous tissues, thereby achieving beauty and anti-aging effects. The effect of non-cross-linked hyaluronic acid gel can generally only last for 4 weeks after one injection, and the first week after injection is the skin recovery period. Therefore, there is an urgent need to improve its shortcomings of short time and poor effect. In order to increase the maintenance time of hyaluronic acid in the body, hyaluronic acid needs to be cross-linked. The degree of cross-linking shows a certain positive correlation with the maintenance time in the body. However, hyaluronic acid with a high degree of cross-linking is not suitable for water-light injection. Reducing the degree of cross-linking of hyaluronic acid can make it moderately soft and hard, and it is not easy to get skin bumps after water-light injection, and it can maintain a longer effective time. However, in the actual production process, in order to ensure the safety of water-light products, they are often required to be sterilized at high temperature. Hyaluronic acid with a low degree of cross-linking is prone to reduced viscosity during sterilization, affecting the effect of water-light injection. Summary of the invention

[0005] In view of the problems existing in the prior art, the present application finds that oligopeptides have the effect of improving the stability of cross-linked hyaluronic acid or its salts, and in particular can improve the stability of the viscosity of cross-linked hyaluronic acid or its salts before and after sterilization.

[0006] In this application, the technical solution is as follows:

[0007] 1. Application of an oligopeptide in improving the stability of cross-linked hyaluronic acid or its salt.

[0008] 2. Use of cross-linked hyaluronic acid or its salt and oligopeptide in the preparation of injectable hyaluronic acid gel.

[0009] 3. According to the use described in item 1 or 2, the oligopeptide has the effect of improving the viscosity stability of cross-linked hyaluronic acid or its salt before and after sterilization.

[0010] 4. According to the use described in item 1 or 2, the oligopeptide is anserine, oligopeptide 1, glutathione or carnosine, preferably anserine or oligopeptide 1.

[0011] 5. The use according to item 1 or 2,

[0012] The cross-linked hyaluronic acid or its salt is (1 to 25) parts by weight, preferably (1.5 to 15) parts by weight, relative to 1 part by weight of the oligopeptide.

[0013] 6. An injectable hyaluronic acid gel, comprising: cross-linked hyaluronic acid or its salt and oligopeptide.

[0014] 7. The injectable hyaluronic acid gel according to item 6,

[0015] The oligopeptide is anserine, oligopeptide 1, glutathione or carnosine, preferably anserine or oligopeptide 1.

[0016] 8. The injectable hyaluronic acid gel according to item 6,

[0017] The cross-linked hyaluronic acid or its salt is (1 to 25) parts by weight, preferably (1.5 to 15) parts by weight, relative to 1 part by weight of the oligopeptide.

[0018] 9. The injectable hyaluronic acid gel according to item 6,

[0019] The gel also includes auxiliary materials and active ingredients.

[0020] 10. According to the injectable hyaluronic acid gel described in item 9, the active ingredients include one or more of vitamins, amino acids, polynucleotides, and polydeoxyribonucleotides.

[0021] Technical Effects

[0022] (1) The viscosity of cross-linked hyaluronic acid decreases after sterilization. Adding oligopeptides at a certain mass ratio can alleviate the decrease in viscosity of cross-linked hyaluronic acid and maintain the stability of cross-linked hyaluronic acid after sterilization.

[0023] (2) Using cross-linked hyaluronic acid gel as the matrix and compounding it with other nutrients can replenish water and moisturize the skin, stimulate the activity of fibroblasts, increase the synthesis of substrates by fibroblasts, and thus improve the quality of the skin.

[0024] (3) The product is terminally sterilized and highly safe for injection. The addition of oligopeptide ingredients, in addition to being a nutrient itself, can protect the viscosity of the cross-linked hyaluronic acid from decreasing during sterilization; it can also stabilize the gel, and the nutrient content and pH value decrease slowly during storage. In particular, when the polypeptide component is combined with polynucleotides or polydeoxyribonucleotides, the protective effect is more obvious. DETAILED DESCRIPTION

[0025] The present application is further described below in conjunction with examples. It should be understood that the examples are only used to further describe and illustrate the present application and are not used to limit the present application.

[0026] Unless otherwise defined, the technical and scientific terms in this specification have the same meaning as those generally understood by those skilled in the art. Although similar or identical methods and materials as described herein may be used in experiments or practical applications, materials and methods are described herein below. In the event of a conflict, the present specification including the definitions therein shall prevail. In addition, materials, methods and examples are provided for illustration only and are not restrictive. The present application is further described below in conjunction with specific embodiments, but is not intended to limit the scope of the present application.

[0027] The present application provides an application of an oligopeptide in improving the stability of cross-linked hyaluronic acid or its salt.

[0028] The present application provides a use of cross-linked hyaluronic acid or its salt and oligopeptide in preparing injectable hyaluronic acid gel.

[0029] In some embodiments of the present application, the oligopeptide is used to improve the stability of the viscosity of cross-linked hyaluronic acid or its salt before and after sterilization.

[0030] In this application, oligopeptide refers to a classification of polypeptides, and the molecular weight range is generally below 1000 Daltons, also known as small peptides, oligopeptides, oligopeptides or small molecule active peptides, compounds formed by the condensation of 2 to 10 amino acids. Those exceeding are called polypeptides, and polypeptides with more than 50 amino acids are called proteins. The difference from other peptides is that it can be directly absorbed in the human body without digestion. Oligopeptides can be divided into: oligopeptide-1, oligopeptide-3, oligopeptide-5, etc., and oligopeptide-6 is also called hexapeptide or hexapeptide.

[0031] In the present application, cross-linked hyaluronic acid or its salt refers to cross-linked hyaluronic acid or its salt obtained by cross-linking hyaluronic acid or its salt with a cross-linking agent. Cross-linked hyaluronic acid or its salt is more stable in molecular structure, lasts longer, and decomposes more slowly.

[0032] Hyaluronic acid or its salt is mixed with a cross-linking agent in an alkaline solution to carry out a cross-linking reaction, and the cross-linking agent is eluted with a buffer solution to terminate the cross-linking reaction, and the cross-linked hyaluronic acid or its salt is obtained after homogenization.

[0033] In the present application, after sterilization, the cross-linked hyaluronic acid or its salt is easily degraded, and the degradation is mainly caused by hydrolysis and active oxygen on the hydroxyl group. Under acidic conditions, hydrolysis occurs at C1, C4 and C connected to the carbonyl C on the uronic acid residue; in alkaline solution, the breakage mostly occurs at C1, O in the ring and N atom on the acetylamino group. The hydrolysis reaction of hyaluronic acid or its salt is caused by random cutting of the molecular chain. The degradation of hyaluronic acid or its salt reduces the viscosity of the solution. When the temperature exceeds 60°C, the viscosity decreases rapidly. The cross-linked hyaluronic acid modified by the cross-linking agent also maintains the characteristics of the hyaluronic acid macromolecule. During wet heat sterilization, the molecular chain breaks, resulting in a significant decrease in viscosity.

[0034] In some embodiments of the present application, the hyaluronate is selected from one or more of the sodium salt, potassium salt, zinc salt, calcium salt, and magnesium salt of hyaluronic acid, preferably the sodium salt of hyaluronic acid.

[0035] In some embodiments of the present application, the hyaluronate is sodium hyaluronate, potassium hyaluronate, magnesium hyaluronate, calcium hyaluronate, zinc hyaluronate, or bismuth hyaluronate.

[0036] In some embodiments of the present application, the oligopeptide is anserine, oligopeptide 1, glutathione or carnosine, preferably anserine or oligopeptide 1.

[0037] In the present application, anserine (β-alanyl-1-methyl-L-histidine) is a highly stable water-soluble dipeptide that relieves hyperuricemia by promoting uric acid excretion. It is naturally present in the skeletal muscle tissue and brain tissue of vertebrates. It has a PK value of about 7.1 and remains intact at low pH values ​​(<3.0).

[0038] In the present application, oligopeptide-1 is a synthetic polypeptide composed of glycine, histidine and lysine. It is a white solid powder that is easily soluble in water. When used in the medical field, it can promote the repair and regeneration of damaged epidermis. As a skin care ingredient, it can increase the synthesis of proteins, especially dermal collagen and elastin, thereby helping the skin to improve metabolism and increase the skin's repair ability.

[0039] In some embodiments of the present application, relative to 1 part by weight of the oligopeptide, the cross-linked hyaluronic acid or its salt is (1 to 25) parts by weight, preferably (1.5 to 15) parts by weight;

[0040] For example, relative to 1 part by weight of the oligopeptide, the cross-linked hyaluronic acid or its salt can be 1 part by weight, 2 parts by weight, 3 parts by weight, 4 parts by weight, 5 parts by weight, 6 parts by weight, 7 parts by weight, 8 parts by weight, 9 parts by weight, 10 parts by weight, 11 parts by weight, 12 parts by weight, 13 parts by weight, 14 parts by weight, 15 parts by weight, 16 parts by weight, 17 parts by weight, 18 parts by weight, 19 parts by weight, 20 parts by weight, 21 parts by weight, 22 parts by weight, 23 parts by weight, 24 parts by weight, 25 parts by weight or any range therebetween.

[0041] The present application provides an injectable hyaluronic acid gel, which comprises: cross-linked hyaluronic acid or its salt and oligopeptide.

[0042] Hydrogel is a gel that uses water as a dispersion medium. It is a cross-linked polymer that can swell in water by introducing a part of hydrophobic groups into a water-soluble polymer with a cross-linked structure. It has high water absorption and high water retention, and has good biocompatibility and water permeability. Injectable hydrogel refers to a hydrogel that can be injected and implanted in the human body. Since it directly enters the human tissue to play the expected role, it must be ensured that the viscoelasticity of the injectable hydrogel meets the expected use, is sterile, and has a certain fluidity for injection.

[0043] Injectable gels can include any suitable biological or synthetic gel. For example, gels can include hyaluronic acid, alginate, cross-linked alginate, collagen, fibrin glue, fibrin clot, poly-(N-isopropylacrylamide), agarose, chitin, chitosan, cellulose, polysaccharides, poly-(oxyalkylenes), copolymers of poly-(oxyethylenes)-poly-(oxypropylenes), poly-(vinyl alcohol), polyacrylates, matrigel or its blends.

[0044] In some specific embodiments, the cross-linked hyaluronic acid or its salt is a hyaluronic acid or its salt with a low degree of cross-linking, for example, slightly cross-linked hyaluronic acid or its salt.

[0045] In the slightly cross-linked hyaluronic acid or its salt, the mass ratio of hyaluronic acid or its salt to the cross-linking agent is 100:(1-2);

[0046] For example, in the slightly cross-linked hyaluronic acid or its salt, the mass ratio of hyaluronic acid or its salt to the cross-linking agent can be 100:1, 100:1.1, 100:1.2, 100:1.3, 100:1.4, 100:1.5, 100:1.6, 100:1.7, 100:1.8, 100:1.9, 100:2 or any range therebetween.

[0047] In some embodiments of the present application, the molecular weight of hyaluronic acid or its salt is 500kDa-3000kDa; for example, the molecular weight of hyaluronic acid or its salt is 500kDa, 600kDa, 700kDa, 800kDa, 900kDa, 1000kDa, 1100kDa, 1200kDa, 1300kDa, 1400kDa, 1500kDa, 1600kDa, 1700kDa, 1800kDa, 1900kDa, 2000kDa, 2100kDa, 2200kDa, 2300kDa, 2400kDa, 2500kDa, 2600kDa, 2700kDa, 2800kDa, 2900kDa, 3000kDa or any range therebetween.

[0048] In some embodiments of the present application, in the cross-linked hyaluronic acid or its salt, the cross-linking agent is carbodiimide, divinyl sulfone, ethylene glycol diglycidyl ether, 1,4-butanediol diglycidyl ether or polyglycerol polyglycidyl ether, preferably 1,4-butanediol diglycidyl ether.

[0049] In some embodiments of the present application, the oligopeptide is anserine, oligopeptide 1, glutathione or carnosine, preferably anserine or oligopeptide 1.

[0050] In some embodiments of the present application, relative to 1 part by weight of the oligopeptide, the cross-linked hyaluronic acid or its salt is (1 to 25) parts by weight, preferably (1.5 to 15) parts by weight;

[0051] For example, relative to 1 part by weight of the oligopeptide, the cross-linked hyaluronic acid or its salt can be 1 part by weight, 2 parts by weight, 3 parts by weight, 4 parts by weight, 5 parts by weight, 6 parts by weight, 7 parts by weight, 8 parts by weight, 9 parts by weight, 10 parts by weight, 11 parts by weight, 12 parts by weight, 13 parts by weight, 14 parts by weight, 15 parts by weight, 16 parts by weight, 17 parts by weight, 18 parts by weight, 19 parts by weight, 20 parts by weight, 21 parts by weight, 22 parts by weight, 23 parts by weight, 24 parts by weight, 25 parts by weight or any range therebetween.

[0052] In some embodiments of the present application, the gel further comprises auxiliary materials and active ingredients.

[0053] In some embodiments of the present application, the gel further includes excipients, which may be pH regulators, osmotic pressure regulators, anesthetics, etc. commonly used in injectable hyaluronic acid gels. The present application does not impose any restrictions, and those skilled in the art may freely choose.

[0054] In some embodiments of the present application, the gel also includes an active ingredient, and the excipient may be a functional ingredient commonly used in injectable hyaluronic acid gels, for example, may include one or more of vitamins, amino acids, polynucleotides, and polydeoxyribonucleotides.

[0055] In some embodiments of the present application, the amino acid is selected from one or more of glycine, proline, hydroxyproline, lysine, isoleucine, leucine, serine, alanine, aspartic acid, tyrosine, glutamic acid, phenylalanine, arginine, valine, threonine, histidine, tryptophan, methionine, cystine and cysteine. More preferably, the amino acid is selected from one or more of alanine, glycine, threonine, arginine, lysine, proline, valine, isoleucine and leucine.

[0056] The term amino acid is intended to refer to naturally occurring amino acids and non-naturally occurring amino acids and amino acid analogs and mimetics. Naturally occurring amino acids include the 20 (L)-amino acids utilized during protein biosynthesis as well as other amino acids, such as 4-hydroxyproline, hydroxylysine, desmosine, isodesmosine, homocysteine, citrulline and ornithine. Non-naturally occurring amino acids include, for example, (D)-amino acids, norleucine, norvaline, p-fluorophenylalanine, ethionine and similar amino acids, which are known to those skilled in the art. Amino acid analogs include modified forms of naturally occurring amino acids and non-naturally occurring amino acids. These modifications may include, for example, substitution or replacement of chemical groups and moieties on the amino acids, or derivatization of the amino acids. Amino acid mimetics include, for example, organic structures that functionally exhibit similar properties, such as charge and charge spacing characteristics of reference amino acids. For example, an organic structure that mimics arginine (Arg or R) will have a positively charged moiety that is located in a similar molecular space and has the same degree of mobility as the ε-amino group of the side chain of the naturally occurring Arg amino acid. Mimetics also include constrained structures to maintain optimal spacing and charge interactions of amino acids or amino acid functional groups. Those skilled in the art know or can determine what structures constitute functionally equivalent amino acid analogs and amino acid mimetics.

[0057] In some embodiments of the present application, the vitamin is selected from one or more of vitamin B1, vitamin B2, niacinamide, vitamin B6, vitamin B12, folic acid, biotin, calcium pantothenate and vitamin C.

[0058] Vitamin B is a water-soluble vitamin. Vitamin B1 (thiamine) can inhibit the activity of cholinesterase. When vitamin B1 is deficient, the activity of this enzyme is too high, and acetylcholine (one of the neurotransmitters) is destroyed in large quantities, affecting nerve conduction, which may affect gastrointestinal motility and the secretion of digestive juices. Thiamine is usually made into a more stable form such as hydrochloride or nitrate for use. Vitamin B2 (riboflavin 1) constitutes many important coenzymes, participates in metabolism and energy release, and helps growth. It can maintain and improve the health of epithelial tissues, such as the mucosal tissue of the digestive tract. Vitamin B3 (niacin) constitutes a coenzyme of dehydrogenase in the body. Vitamin B3 is the most needed vitamin in the B vitamins. It is not only a vitamin that maintains the health of the digestive system, but also can relieve gastrointestinal disorders. Niacin is converted into nicotinamide in the human body. Nicotinamide is a component of coenzyme I and coenzyme II, and participates in lipid metabolism, the oxidation process of tissue respiration and the anaerobic decomposition of carbohydrates in the body. Vitamin B6 includes pyridoxine, pyridoxal and pyridoxamine, which can be converted into each other. Deoxyadenosylcobalamin is the main form of vitamin B12 in the body. Its role is often related to that of folic acid. Folic acid has a variety of coenzyme forms in cells. The side chain carboxyl group of biotin can be connected to the lysine residue of the enzyme through an amide bond. Biotin is a carboxyl carrier and is also involved in the metabolism of vitamin B12, folic acid and pantothenic acid.

[0059] "Riboflavin" is used interchangeably with "vitamin B2" and includes riboflavin and its esters, particularly riboflavin-5'-phosphate.

[0060] Vitamin C is water-soluble (its aqueous solution is acidic) and is a very strong antioxidant. It can also protect other antioxidants, such as vitamin A, vitamin E, unsaturated fatty acids, and defend free radicals from harm to the human body. The strength of the blood vessel wall has a great relationship with vitamin C. Capillaries are the smallest of all blood vessels. The wall may only have the thickness of a cell. Its strength and elasticity are determined by the collagen responsible for connecting cells with the effect of cement. The presence of vitamin C helps the skin to maintain elasticity and vitality. Ascorbic acid is interchangeable with vitamin C, and also includes its pharmaceutically acceptable salts (such as sodium ascorbate and calcium ascorbate) and its pharmaceutically acceptable esters (particularly palmitic acid ascorbyl ester).

[0061] In some embodiments of the present application, the gel further comprises polynucleotides and / or polydeoxyribonucleotides.

[0062] In some embodiments of the present application, the injectable hyaluronic acid gel is prepared by sterilizing a composite gel including the cross-linked hyaluronic acid or its salt and the oligopeptide.

[0063] In some embodiments of the present application, the cross-linked hyaluronic acid or its salt gel is prepared by mixing hyaluronic acid or its salt with a cross-linking agent in an alkaline solution to carry out a cross-linking reaction, eluting the cross-linking agent with a buffer solution to terminate the cross-linking reaction, and obtaining a cross-linked hyaluronic acid or its salt gel after homogenization; mixing: uniformly mixing the cross-linked hyaluronic acid or its salt gel with a nutrient solution containing oligopeptides to obtain a composite gel; sterilization: canning the composite gel and sterilizing it with wet heat to obtain an injectable hyaluronic acid or its salt composite gel.

[0064] In some embodiments of the present application, the cross-linked hyaluronic acid or its salt gel is prepared by: mixing hyaluronic acid or its salt with a cross-linking agent in an alkaline solution to carry out a cross-linking reaction, eluting the cross-linking agent with a buffer solution to terminate the cross-linking reaction, and obtaining a cross-linked hyaluronic acid or its salt gel after homogenization; mixing: uniformly mixing the cross-linked hyaluronic acid or its salt gel with a solution containing oligopeptides, vitamins, amino acids, polynucleotides and / or polydeoxyribonucleotides to obtain a composite gel; sterilization: canning the composite gel and sterilizing it with wet heat to obtain an injectable hyaluronic acid or its salt composite gel.

[0065] Since injectable hyaluronic acid gel is directly injected into the human body, it needs to be sterilized during the preparation process. After sterilization, the cross-linked hyaluronic acid or its salt is prone to degradation. The degradation is mainly caused by hydrolysis and active oxygen on the hydroxyl group. Under acidic conditions, hydrolysis occurs at C1, C4 and C connected to the carbonyl C on the uronic acid residue; in alkaline solution, the cleavage mostly occurs at C1, O in the ring and N atom on the acetylamino group. The hydrolysis reaction of hyaluronic acid or its salt is caused by random cutting of the molecular chain. The degradation of hyaluronic acid or its salt reduces the viscosity of the solution. When the temperature exceeds 60°C, the viscosity decreases rapidly. The cross-linked hyaluronic acid modified by the cross-linking agent also maintains the characteristics of the hyaluronic acid macromolecule. During wet heat sterilization, the molecular chain breaks, resulting in a significant decrease in viscosity. In the injectable hyaluronic acid gel, the addition of oligopeptides can not only serve as a nutrient itself, but also protect the viscosity of the cross-linked hyaluronic acid from decreasing during sterilization; it can also stabilize the gel, and the nutrient content decreases slowly during storage.

[0066] Example 1

[0067] Take 0.01g of 1,4-butanediol diglycidyl ether (BDDE) and mix it evenly with 5mL of 1wt% sodium hydroxide solution, then add 1.0g of sodium hyaluronate and stir to dissolve it evenly. Seal the above reactants, let them stand at 3℃ for 15h, then let them stand at 50℃ for 3h, and finally let them stand at 20℃ for 15h. The reactants are dialyzed, swollen and homogenized in phosphate sodium chloride buffer to obtain a cross-linked sodium hyaluronate gel with a content of 2mg / mL; add an anserine solution with a content of 2mg / mL to the cross-linked sodium hyaluronate gel at a volume ratio of 1:1, mix evenly, and obtain a cross-linked sodium hyaluronate gel, wherein the content of the cross-linked sodium hyaluronate gel is 1mg / mL, the content of anserine is 1mg / mL, and the mass ratio of oligopeptide to cross-linked sodium hyaluronate is 1:1. Determine the viscosity of the gel before and after sterilization.

[0068] Viscosity determination method: Haake RS6000 (Thermo Fisher Scientific (China) Co., Ltd.) was used for determination under the following conditions: rotor: P20TiL; Gap value: 1.00 mm; temperature: 25°C; determination mode: oscillation frequency sweep CD; stress: 1%; frequency range: 0.01-1 Hz. The viscosity value at 0.1 Hz was recorded.

[0069] Example 2

[0070] The only difference between Example 2 and Example 1 is that the content of the cross-linked sodium hyaluronate gel is 1 mg / mL, the content of anserine is 0.5 mg / mL, the mass ratio of the oligopeptide to the cross-linked sodium hyaluronate is 1:2, and the other conditions are the same.

[0071] Example 3

[0072] The only difference between Example 3 and Example 1 is that the content of the cross-linked sodium hyaluronate gel is 1 mg / mL, the content of anserine is 0.1 mg / mL, the mass ratio of the oligopeptide to the cross-linked sodium hyaluronate is 1:10, and the other conditions are the same.

[0073] Example 4

[0074] The only difference between Example 4 and Example 1 is that the content of the cross-linked sodium hyaluronate gel is 1 mg / mL, the content of anserine is 0.04 mg / mL, the mass ratio of the oligopeptide to the cross-linked sodium hyaluronate is 1:25, and the other conditions are the same.

[0075] Example 5

[0076] The only difference between Example 5 and Example 1 is that the oligopeptide is oligopeptide 1, the content of the cross-linked sodium hyaluronate gel is 1 mg / mL, the content of oligopeptide 1 is 1 mg / mL, the mass ratio of the oligopeptide to the cross-linked sodium hyaluronate is 1:1, and the other conditions are the same.

[0077] Example 6

[0078] The only difference between Example 6 and Example 5 is that the content of the cross-linked sodium hyaluronate gel is 1 mg / mL, the content of oligopeptide 1 is 0.5 mg / mL, the mass ratio of oligopeptide to cross-linked sodium hyaluronate is 1:2, and the other conditions are the same.

[0079] Example 7

[0080] The only difference between Example 7 and Example 5 is that the content of the cross-linked sodium hyaluronate gel is 1 mg / mL, the content of oligopeptide 1 is 0.1 mg / mL, the mass ratio of oligopeptide to cross-linked sodium hyaluronate is 1:10, and the other conditions are the same.

[0081] Example 8

[0082] The only difference between Example 8 and Example 5 is that the content of the cross-linked sodium hyaluronate gel is 1 mg / mL, the content of oligopeptide 1 is 0.04 mg / mL, the mass ratio of oligopeptide to cross-linked sodium hyaluronate is 1:25, and the other conditions are the same.

[0083] Example 9

[0084] The only difference between Example 9 and Example 1 is that the oligopeptide is carnosine, the content of the cross-linked sodium hyaluronate gel is 1 mg / mL, the content of carnosine is 0.1 mg / mL, the mass ratio of the oligopeptide to the cross-linked sodium hyaluronate is 1:10, and the other conditions are the same.

[0085] Example 10

[0086] The only difference between Example 10 and Example 1 is that the oligopeptide is glutathione, the content of the cross-linked sodium hyaluronate gel is 1 mg / mL, the content of glutathione is 0.1 mg / mL, the mass ratio of the oligopeptide to the cross-linked sodium hyaluronate is 1:10, and the other conditions are the same.

[0087] Comparative Example 1

[0088] The only difference between Comparative Example 1 and Example 1 is that the oligopeptide is not contained, the content of the cross-linked sodium hyaluronate gel is 1 mg / mL, and the other conditions are the same.

[0089] Table 1

[0090]

[0091]

[0092] As can be seen from Table 1, the viscosity of cross-linked hyaluronic acid alone changes greatly before and after sterilization. The addition of oligopeptides can reduce the viscosity change of cross-linked hyaluronic acid before and after sterilization. It can be seen from Examples 1-8 that whether it is anserine or oligopeptide 1, when the mass ratio of oligopeptide to cross-linked sodium hyaluronate is 1: (1.5-15), the viscosity of cross-linked sodium hyaluronate gel decreases to the lowest degree and has the best stability. When the mass ratio of oligopeptide to cross-linked sodium hyaluronate is 1: 1 or 1: 25, the viscosity of cross-linked sodium hyaluronate gel decreases to a low degree and has good stability. When the oligopeptide is replaced by carnosine or glutathione, the viscosity of cross-linked sodium hyaluronate gel decreases to a low degree and has good stability. When no oligopeptide is added, the viscosity of cross-linked sodium hyaluronate gel decreases seriously.

[0093] Table 2 Composite gel

[0094]

[0095] Embodiment 11

[0096] Take 0.01 g of BDDE and 5 mL of 1 wt% sodium hydroxide solution and mix them evenly, then add 1.0 g of sodium hyaluronate and stir to make it evenly dissolved. Seal the above reactants, place them at 3°C ​​for 15 hours, then place them at 50°C for 3 hours, and finally place them at 20°C for 15 hours. The final reaction product is dialyzed, swollen, and homogenized in a phosphate sodium chloride buffer to obtain a cross-linked sodium hyaluronate gel with a content of 16 mg / mL; a composite solution is added to the cross-linked sodium hyaluronate gel at a volume ratio of 1:1, wherein the content of anserine in the composite solution is 2 mg / mL, the content of nicotinamide is 0.04 mg / mL, the content of alanine is 0.4 mg / mL, the content of polynucleotide is 1.0 mg / mL, and the content of polydeoxyribonucleotide is 20 mg / mL, and the composite gel is obtained by mixing evenly. In the composite gel shown, the content of cross-linked sodium hyaluronate gel is 8 mg / mL, the content of anserine is 1 mg / mL, the content of nicotinamide is 0.02 mg / mL, the content of alanine is 0.2 mg / mL, the content of polynucleotide is 0.5 mg / mL, and the content of polydeoxyribonucleotide is 10 mg / mL. The composite gel is canned and sterilized by wet heat to obtain an injectable hyaluronic acid or its salt composite gel.

[0097] Comparative Example 2

[0098] The only difference between Comparative Example 2 and Example 11 is that it does not contain oligopeptides, the content of cross-linked sodium hyaluronate gel is 8 mg / mL, the content of anserine is 1 mg / mL, the content of nicotinamide is 0.02 mg / mL, the content of alanine is 0.2 mg / mL, the content of polynucleotide is 0.5 mg / mL, the content of polydeoxyribonucleotide is 10 mg / mL, and the other conditions are the same.

[0099] The viscosity of Example 11 and Comparative Example 2 before and after sterilization was measured by referring to the method of Example 1. The viscosity change of Example 11 before and after sterilization was 17.2%, and the viscosity change of Comparative Example 2 before and after sterilization was 61.8%, that is, the viscosity of the injectable hyaluronic acid or its salt composite gel was significantly reduced after the addition of oligopeptides.

[0100] Although the present invention has been disclosed as above by way of embodiments, it is not intended to limit the present invention. Any person having ordinary knowledge in the technical field may make some changes and modifications without departing from the spirit and scope of the present invention. Therefore, the protection scope of the present invention shall be determined by the scope of the patent application attached hereto.

Claims

1. Use of an oligopeptide in improving the stability of cross-linked hyaluronic acid or its salt; The oligopeptide has the effect of improving the viscosity stability of cross-linked hyaluronic acid or its salt before and after moist heat sterilization; the oligopeptide is anserine, oligopeptide 1, glutathione or carnosine; relative to 1 part by weight of the oligopeptide, the cross-linked hyaluronic acid or its salt is 1 to 25 parts by weight; The cross-linked hyaluronic acid or its salt gel is evenly mixed with the nutrient solution containing oligopeptides to obtain a composite gel; Sterilization: the composite gel is canned and sterilized by wet heat.

2. The use according to claim 1, characterized in that: The oligopeptide is anserine or oligopeptide 1.

3. The use according to claim 1 or 2, characterized in that: The cross-linked hyaluronic acid or its salt is 1.5 to 15 parts by weight relative to 1 part by weight of the oligopeptide.

4. An injectable hyaluronic acid gel comprising: Cross-linked hyaluronic acid or its salt and oligopeptide; the oligopeptide is anserine or oligopeptide 1, and the cross-linked hyaluronic acid or its salt is 1.5 to 15 parts by weight relative to 1 part by weight of the oligopeptide; Evenly mixing the cross-linked hyaluronic acid or its salt gel with the nutrient solution containing oligopeptides to obtain a composite gel; Sterilization: The composite gel is canned and sterilized by wet heat to obtain an injectable hyaluronic acid gel.

5. The injectable hyaluronic acid gel according to claim 4, characterized in that The gel also includes auxiliary materials and active ingredients.

6. The injectable hyaluronic acid gel according to claim 5, characterized in that The active ingredients include one or more of vitamins, amino acids, polynucleotides, and polydeoxyribonucleotides.

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