PEG (Polyethylene Glycol) derivative cross-linked hyaluronic acid polymer as well as preparation method and application thereof

Abstract

The invention provides a PEG derivative cross-linked hyaluronic acid polymer as well as a preparation method and application thereof. The polymer is formed by cross-linking hyaluronic acid or a hyaluronic acid derivative and a PEG derivative, and has a structure as shown in formula I: A is provided by the PEG derivative, Y is a linking group, G is provided by hyaluronic acid or a derivative thereof, and m is greater than or equal to 1 and less than or equal to the number of functional groups of the PEG derivative. The polymer is high in biological safety and good in viscoelasticity, a cross-linked network can be sterilized through high-temperature steam, finally stably exists in a sterile form, and the polymer can be used for industrialization and has high application value.

Description

A PEG derivative cross-linked hyaluronic acid polymer and its preparation method and application

[0001] This application claims priority to the prior application filed by the applicant with the State Intellectual Property Office of China on September 5, 2023, with patent application number 202311138732.2, and invention name “A PEG derivative cross-linked hyaluronic acid polymer, its preparation method and application”. Technical Field

[0002] The present invention belongs to the field of polymers, and in particular relates to a PEG derivative cross-linked hyaluronic acid polymer and a preparation method and application thereof. Background Art

[0003] Dermal fillers, or soft tissue fillers, are widely used in plastic surgery for cosmetic applications or tissue reconstruction. Hyaluronic acid (HA) is widely used as a dermal filler due to its biodegradability, biocompatibility, and non-immunogenicity. However, due to the presence of hyaluronidase in the body, free HA has a short half-life in vivo and lacks tissue support, necessitating chemical modification with cross-linking agents to achieve the desired properties. The vast majority of cross-linked sodium hyaluronate products currently on the market use toxic cross-linking agents, such as 1,4-butanediol diglycidyl ether (BDDE) (Juvéderm, Restylane, Princess), divinyl sulfone (DVS) (Captique, Hylaform, Prevelle), or diepoxyoctane (Puragen), such as Juvéderm ULTRA (National Medical Device Registration No. 20153131709) produced by Allergan and Restylane Volyme (National Medical Device Registration No. 20233130011) produced by Q-Med AB. Literature reports indicate that these commonly used crosslinkers are biotoxic. For example, BDDE-crosslinked HA gels at a concentration of 0.5 mg / mL induce morphological changes and reduced cell viability in mouse fibroblasts, while DVS-crosslinked HA at a concentration of 100 mM reduces retinal pigment epithelial cell viability and increases the expression of inflammatory cytokines. Although manufacturers have set limits on the residual amounts of various crosslinkers, such as limiting BDDE residues to no more than 2.0 μg / g, multiple or high-dose fillings can pose safety risks and significantly increase the incidence of various inflammatory reactions or adverse reactions. Therefore, the development of safe and non-toxic crosslinkers and crosslinking processes is imperative.

[0004] Sodium hyaluronate is a mucopolysaccharide composed of N-acetylglucosamine and D-glucose linked by β(1-4) glycosidic bonds. Sodium hyaluronate contains four sites amenable to chemical modification: carboxyl, hydroxyl, acetylamino, and reducing ends. Therefore, sodium hyaluronate can be cross-linked with other polymers through various cross-linking bonds, thereby achieving a three-dimensional network structure that provides support.

[0005] In recent years, with the gradual development of synthetic chemistry, the number of functional groups, type of functional groups, molecular weight, and purity of multifunctional (functional group number ≥ 2) PEG (polyethylene glycol) derivatives can be well controlled. PEG derivatives also have good biocompatibility, so multifunctional PEG derivatives are widely used in pharmaceutical research and development. They are currently mainly used to modify protein drugs to increase their in vivo half-life, reduce immunogenicity, and increase the drug's water solubility. Currently, Swiss company MatexLab Spa uses polyethylene glycol diglycidyl ether (PEGDE) as a crosslinker to develop HA implant products. However, the epoxy group of PEGDE is similar to BDDE and still has biotoxicity. Therefore, a new type of non-biotoxic hydrogel material is needed to address the problems in these applications.

[0006] Summary of the Invention

[0007] In order to improve the above technical problems, the present invention provides a polymer, which is formed by cross-linking hyaluronic acid or a hyaluronic acid derivative with a PEG derivative, and has a structure as shown in Formula I:

[0008] A is provided by a PEG derivative, wherein the PEG derivative is selected from a multi-arm PEG terminated with a functional group, wherein the functional group is selected from at least one of a hydroxyl group, an amino group, and a carboxyl group, and the multi-arm refers to no less than two arms, for example, at least one selected from two to sixteen arms, etc.; for example, the PEG derivative can be selected from at least one of a two-arm PEG, a three-arm PEG, a four-arm PEG, a five-arm PEG, a six-arm PEG, a seven-arm PEG, an eight-arm PEG, a nine-arm PEG, a ten-arm PEG, an eleven-arm PEG, a twelve-arm PEG, a thirteen-arm PEG, a fourteen-arm PEG, a fifteen-arm PEG, a sixteen-arm PEG, etc. terminated with a hydroxyl group, an amino group, and / or a carboxyl group; preferably, the PEG derivative is selected from a two-arm PEG, a four-arm PEG, or an eight-arm PEG terminated with a hydroxyl group, an amino group, and / or a carboxyl group; further preferably, the PEG derivative can be an eight-arm PEG terminated with a hydroxyl group, an eight-arm PEG terminated with a carboxyl group, an amino-terminated two-arm PEG, an amino-terminated eight-arm PEG, or an eight-arm PEG terminated with both a hydroxyl group and an amino group;

[0009] Y is a linking group selected from a strongly polar group, preferably at least one of an ester group and an amide group;

[0010] G is provided by hyaluronic acid or a derivative thereof; for example, the weight average molecular weight of the hyaluronic acid or a derivative thereof is 1 million Daltons to 5 million Da, preferably 1.5 million Da to 5 million Da; further preferably, the weight average molecular weight of the hyaluronic acid or a derivative thereof is 180 Da to 3 million Da;

[0011] 1≤m≤the number of functional groups of the PEG derivative, preferably m is an integer, for example, m is 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16.

[0012] According to an embodiment of the present invention, the molecular weight of the PEG derivative is 100-100000 Da, for example, 300-80000 Da, preferably 500-50000 Da, more preferably 1000-20000 Da, exemplified by 200 Da, 300 Da, 400 Da, 600 Da, 700 Da, 800 Da, 900 Da, 1100 Da, 1200 Da, 1300 Da, 1400 Da, 1500 Da, 1600 Da, 1700 Da, 1800 Da, 1900 Da, 2000 Da, 4000 Da, 5000 Da, 7000 Da, 10000 Da, and 15000 Da.

[0013] According to an embodiment of the present invention, G is provided by sodium hyaluronate. Preferably, the weight average molecular weight of the sodium hyaluronate is 1.35 million Da, 1.4 million Da, 1.5 million Da, 1.6 million Da, 1.7 million Da, 1.8 million Da, 1.9 million Da, 2 million Da, 2.1 million Da, 2.2 million Da, 2.3 million Da, 2.4 million Da, 2.5 million Da, 2.6 million Da, 2.7 million Da, 2.8 million Da, 2.9 million Da, 3.5 million Da, 4 million Da or 4.5 million Da.

[0014] According to an embodiment of the present invention, the degree of polymerization x of the polymer may be selected from 1000-8000, such as 2000-6000.

[0015] According to an embodiment of the present invention, the polymer is formed by cross-linking sodium hyaluronate with a PEG derivative.

[0016] In Formula I, A is provided by a PEG derivative, wherein the PEG derivative is selected from a hydroxyl-terminated eight-arm PEG, a carboxyl-terminated eight-arm PEG, an amino-terminated two-arm PEG, an amino-terminated eight-arm PEG, or an eight-arm PEG terminated with both a hydroxyl group and an amino group, and the molecular weight of the PEG derivative is 1000-20000 Da;

[0017] Y is a linking group selected from an ester group and / or an amide group;

[0018] G is provided by sodium hyaluronate, and the weight average molecular weight of the sodium hyaluronate is 1.8-3 million Da.

[0019] The present invention also provides a method for preparing the above-mentioned polymer, comprising the following steps: mixing a PEG derivative, hyaluronic acid or a derivative thereof, a condensing agent and a buffer, homogenizing the mixture to obtain a uniform solution, and performing a cross-linking reaction to obtain the polymer;

[0020] The PEG derivatives, hyaluronic acid or derivatives thereof have the definitions shown above, respectively.

[0021] According to an embodiment of the present invention, the condensing agent can be selected from at least one of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC), 4-(4,6-dimethoxytriazine-2-yl)-4-methylmorpholine hydrochloride (DMTMM), N-hydroxysuccinimide (NHS) and N-hydroxysulfosuccinimide.

[0022] According to an embodiment of the present invention, the mass concentration of hyaluronic acid or its derivatives in the mixture is 0.5%-4%, preferably 1%-3%, for example 0.75%, 1.2%, 1.5%, 2%, 2.5%, 3.5%.

[0023] According to an embodiment of the present invention, the mass concentration of the PEG derivative in the mixture is 0.05%-1%, preferably 0.25%-0.8%, for example 0.06%, 0.07%, 0.075%, 0.08%, 0.09%, 0.15%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.9%.

[0024] According to an embodiment of the present invention, the mass concentration of the condensing agent in the mixture is 0.005%-0.6%, preferably 0.2%-0.4%, such as 0.01%, 0.05%, 0.1%, 0.3%, 0.5%.

[0025] According to an embodiment of the present invention, the homogenization treatment time is 0.5h-6h, preferably 2-4h.

[0026] According to an embodiment of the present invention, the buffer solution may be a phosphate buffer solution or a (N-morpholino)ethanesulfonic acid buffer solution.

[0027] According to an embodiment of the present invention, the temperature of the cross-linking reaction is 4°C-35°C, preferably 15°C-28°C.

[0028] According to an embodiment of the present invention, the cross-linking reaction time is 36h-120h, preferably 60h-96h.

[0029] According to an embodiment of the present invention, the preparation method further comprises dialyzing the product after the cross-linking reaction is completed. For example, the dialysis temperature is 4°C-35°C, preferably 15°C-28°C.

[0030] According to an embodiment of the present invention, the dialysis can be performed on a shaker, and the shaker speed can be 40 rpm-80 rpm, preferably 50 rpm-70 rpm.

[0031] According to an embodiment of the present invention, the preparation method further comprises sterilization: sterilizing the final product obtained by dialysis to obtain a sterilized product.

[0032] According to an embodiment of the present invention, the sterilization includes high-temperature steam sterilization, for example, the sterilization temperature is 121 degrees Celsius and the sterilization time is 15 minutes.

[0033] The preparation principle of the polymer of the present invention is as follows:

[0034] 1. Esterification reaction cross-linked hydrogel: 1) Activating the carboxyl groups of the carboxyl-terminated multi-arm PEG derivative with a condensing agent to couple the carboxyl groups with the hydroxyl groups on the sodium hyaluronate molecular chain; 2) Activating the carboxyl groups on the sodium hyaluronate molecular chain with a condensing agent to couple the carboxyl groups with the hydroxyl groups on the hydroxyl-terminated multi-arm PEG derivative; then, removing the condensing agent by washing; and finally, sterilizing the hydrogel with high-temperature steam to obtain the corresponding esterification reaction cross-linked polymer;

[0035] 2. Amidation reaction cross-linked hydrogel: A condensing agent is used to activate the carboxyl groups on the sodium hyaluronate molecular chain, allowing it to couple with the amino groups of the amino-terminated multi-arm PEG derivative. The condensing agent is then removed by washing, and finally sterilized by high-temperature steam to obtain the corresponding amidation reaction cross-linked polymer.

[0036] 3. Esterification reaction combined with amidation reaction to form a double-crosslinked hydrogel: A condensing agent is used to activate the carboxyl groups on the sodium hyaluronate molecular chain and the multi-arm PEG derivatives with carboxyl or hydroxyl and amino groups, so that they are coupled with the hydroxyl and amino groups on the molecular chains in the reaction system. The condensing agent is then removed by washing, and finally sterilized by high-temperature steam to obtain the corresponding double-crosslinked polymer.

[0037] The present invention also provides a gel comprising the above polymer and a liquid, wherein the polymer has a cross-linked network structure, and the liquid is stored in the network structure. Preferably, the liquid is water, that is, the gel is a hydrogel.

[0038] The present invention further provides a composition comprising the above polymer or the gel.

[0039] According to an embodiment of the present invention, the composition may further comprise an auxiliary material.

[0040] According to an embodiment of the present invention, the composition may further comprise an additive, for example, the additive may be selected from at least one of a preservative, a solubilizer, a surfactant, a buffer, an isotonicity regulator, a suspending agent, a dispersant, a wetting agent, and the like.

[0041] The present invention further provides a preparation containing the above polymer, gel or composition.

[0042] According to an embodiment of the present invention, the preparation can be any one of tablets, capsules, powders, granules, syrups, gels, suspensions, dispersible tablets, chewable tablets, effervescent tablets, capsules, granules, sustained-release preparations, injections, transdermal absorption preparations, etc.

[0043] According to an embodiment of the present invention, the active ingredient of the preparation includes the above composition, or further includes a double-crosslinked polysaccharide polymer.

[0044] The present invention further provides the use of the polymer, the gel or the composition in preparing products, and the products can be medicines, pharmaceutical excipients, medical devices, cosmetics, cosmetic raw materials, skin care products, skin care raw materials, women's care products, daily necessities, foods, food additives, dietary supplements, health products, meal replacements, dietary supplements, veterinary medicines, veterinary foods, veterinary medical devices, veterinary care products, etc., and can also be industrial products.

[0045] The present invention further provides the use of the polymer, the gel or the composition in the preparation of any product for tissue regeneration, tissue repair, tissue filling, wound care, tissue adhesion prevention, postoperative care, wrinkle removal, plastic surgery, medical cosmetology, daily beauty, anti-aging, whitening skin care, scalp care, hair transplantation, hair growth, fever reduction, lubrication, etc.

[0046] The present invention further provides use of the polymer, the gel or the composition in preparing medicines for preventing and / or treating diseases related to overweight.

[0047] The present invention further provides a method for preventing and / or treating diseases associated with excess weight, comprising administering a suitable dose of the above polymer, gel, composition or preparation to a subject in need thereof.

[0048] In some specific embodiments, the diseases associated with overweight may include at least one of overweight, obesity, prediabetes, diabetes, hypercholesterolemia, hypertension, cardiovascular and cerebrovascular diseases, constipation, osteoarthritis, gout, sleep apnea syndrome, infertility, menstrual disorders, abnormal ovulation, abnormal hormone levels, etc.

[0049] In some embodiments, a suitable dosage may comprise a predetermined amount of at least one of the polymer, the gel, or the composition calculated to produce the desired preventive, therapeutic, or ameliorative effect in combination with a desired carrier. Medical professionals skilled in the art can determine the therapeutically effective amount based on patient characteristics well known in the art, such as age, weight, sex, symptoms, complications, other diseases, and the like.

[0050] The term "at least one" means one, two or more. Beneficial effects

[0051] The polymer provided by the present invention, namely, PEG derivative cross-linked hyaluronic acid, does not add toxic cross-linking agents during the preparation process. Only multi-arm PEG derivatives blocked with hydroxyl, amino, carboxyl, etc. are used as cross-linking agents, and are directly cross-linked with hyaluronic acid or its derivatives through esterification reaction or amidation reaction. The obtained polymer has high biological safety and good viscoelasticity. The cross-linked network can be sterilized by high-temperature steam and ultimately exists in a sterile and stable form. It can be used for industrialization and has high application value. DETAILED DESCRIPTION

[0052] The polymer mentioned above is formed by cross-linking hyaluronic acid or a hyaluronic acid derivative with a PEG derivative, and has a repeating unit structure as shown in Formula I:

[0053] The degree of polymerization x of the polymer is 1000-8000, for example 2000-5000, such as 3000, 3500, 4000, 4500, 5500, 6000.

[0054] In one embodiment, the polymer has a structure as shown in Formula II or Formula III:

[0055] Wherein, Formula II is a schematic structural diagram of an eight-arm PEG in which A is a hydroxyl or carboxyl-terminated group and m is 4;

[0056] Formula III is a schematic structural diagram when A is an amine-terminated two-arm PEG and m is 2;

[0057] n, x are the degree of polymerization, which meet the actual molecular weight required for the polymer. In specific common examples, n in the A structure can be selected from 2-800, n in the G structure can be selected from 1000-20000, and x of the polymer can be selected from 1000-8000. It should be noted that the cross-linking reaction sites of the polymer of the present invention are not fixed, and the degree of polymerization does not need to be accurately controlled. Therefore, Formula II, Formula III and m, n, and x therein are only structural schematics and cannot be regarded as limitations on the actual structure of the actual product.

[0058] The technical solutions of the present invention will be described in further detail below with reference to specific embodiments. It should be understood that the following embodiments are merely illustrative and explanations of the present invention and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are encompassed within the scope of protection that the present invention is intended to protect.

[0059] Unless otherwise specified, the raw materials and reagents used in the following examples are commercially available or can be prepared by known methods.

[0060] Examples 1-3

[0061] Examples 1-3 describe the preparation process of the cross-linking reaction between sodium hyaluronate and 8arm-PEG-OH, and detect the elastic modulus and viscous modulus of the final product.

[0062] 1.12 g of sodium hyaluronate of different molecular weights, 0.35 g of hydroxyl-terminated eight-arm polyethylene glycol (8arm-PEG-OH) with a molecular weight of 10,000 Da, and 0.2 g of DMTMM were dissolved in 40 mL of phosphate buffer solution. The weight-average molecular weights of the sodium hyaluronate were 1.35 million Da (Example 1), 2 million Da (Example 2), and 2.8 million Da (Example 3), respectively.

[0063] 2. Use a homogenizer to homogenize for 2 hours to dissolve and homogenize the raw materials in step 1.

[0064] 3. The homogeneous solution obtained in step 2 was placed in an oven at 20°C for cross-linking for 72 hours to obtain a solid gel.

[0065] 4. Place the solid gel in step 3 in phosphate buffer solution and dialyze in a shaker at 20°C with a shaker speed of 40 rpm.

[0066] 5. The gel obtained in step 4 was placed in a high-temperature steam autoclave at 121°C for 15 minutes to obtain sterile 8-arm-PEG-OH cross-linked sodium hyaluronate, designated as Sample 1, Sample 2, and Sample 3. The structures of Samples 1-3 are shown in Formula II, and the degree of polymerization (x) of the polymer ranged from 2000 to 5000.

[0067] 6. Samples 1, 2, and 3 obtained in step 5 were taken for rheological property testing to study the elastic modulus and viscous modulus. The results are shown in Table 1. Table 1 shows that the prepared gel still maintains very high viscoelasticity after sterilization, wherein the elastic modulus and viscous modulus of sample 2 (sodium hyaluronate gel with a molecular weight of 2 million Da) at 5 Hz are 392.4 Pa and 206.5 Pa, respectively, and the elastic modulus and viscous modulus of sample 3 (sodium hyaluronate gel with a molecular weight of 2.8 million Da) at 5 Hz are 259.3 Pa and 133.4 Pa, respectively. The elastic modulus and viscous modulus of samples 2 and 3 at 5 Hz are both greater than those of the commercially available product Juvéderm ULTRA cross-linked sodium hyaluronate gel for injection (Allergan, registration certificate number: Guoxie Zhujin 20153131709, elastic modulus and viscous modulus at 5 Hz are 195 Pa and 74 Pa, respectively). Juvéderm ULTRA injectable cross-linked sodium hyaluronate gel is made by cross-linking 24 mg / mL sodium hyaluronate in phosphate buffer solution with the cross-linking agent BDDE, which is toxic.

[0068] Table 1 Elastic modulus and viscous modulus of HA-8arm-PEG-OH gel after cross-linking with sodium hyaluronate of different molecular weights

[0069] Examples 4-7

[0070] 1. Different masses of amino-terminated two-arm polyethylene glycol (2arm-PEG-NH2) with a molecular weight of 1000 Da, 0.8 g of sodium hyaluronate with a weight-average molecular weight of 1.9 million Da, 0.10 g of EDC, and 0.1 g of NHS were dissolved in 40 mL of phosphate buffer solution, where the masses of 2arm-PEG-NH2 were 0.05 g, 0.1 g, 0.2 g, and 0.4 g, and the corresponding concentrations were 0.125% (Example 4), 0.25% (Example 5), 0.5% (Example 6), and 1% (Example 7), respectively.

[0071] 2. Use a homogenizer to homogenize for 3 hours to dissolve and homogenize the raw materials.

[0072] 3. The homogeneous solution obtained in step 2 was placed in an oven at 25°C for cross-linking for 65 h to obtain a solid gel.

[0073] 4. Place the solid gel in step 3 in phosphate buffer solution and dialyze in a shaker at 25°C with a shaker speed of 60 rpm.

[0074] 5. Place the gel obtained in step 4 in a high-temperature steam autoclave at 121°C for 15 minutes to obtain sterile 2arm-PEG-NH2 cross-linked sodium hyaluronate. The structures of Examples 4-7 can be referred to as Formula III, where x ranges from 2000 to 4500.

[0075] 6. The gel obtained in step 5 was subjected to rheological property testing to study the elastic modulus and viscous modulus. The results are shown in Table 2. As the content of the cross-linking agent 2arm-PEG-NH2 increased, the elastic modulus of the gel gradually increased to 299.4 Pa. Subsequently, as the content of the cross-linking agent was further increased, the elastic modulus gradually decreased to 254.1 Pa. Therefore, the rheological properties of the gel can be controlled by adjusting the content of the cross-linking agent.

[0076] Table 2 Elastic modulus and viscous modulus of gels cross-linked with different concentrations of 2arm-PEG-NH2

[0077] Example 8

[0078] 1. Dissolve 0.96 g of sodium hyaluronate with a weight-average molecular weight of 2.8 million Da, 0.35 g of carboxyl-terminated eight-arm polyethylene glycol (8arm-PEG-COOH) with a molecular weight of 2000 Da, 0.08 g of EDC, and 0.08 g of NHS in 40 mL of phosphate buffer solution.

[0079] 2. Use a homogenizer to homogenize for 3 hours to dissolve and homogenize the raw materials.

[0080] 3. The homogeneous solution obtained in step 2 was placed in an oven at 25°C for cross-linking for 65 h to obtain a solid gel.

[0081] 4. Place the solid gel in step 3 in phosphate buffer solution and dialyze in a shaker at 25°C with a shaker speed of 60 rpm.

[0082] 5. Place the gel obtained in step 4 in a high-temperature steam autoclave at 121°C for 15 minutes to obtain sterile 8-arm-PEG-COOH cross-linked sodium hyaluronate. Its structure is shown in Formula II, where x ranges from 3000 to 4500.

[0083] The eight-arm polyethylene glycol cross-linked hyaluronic acid prepared by the above method has high viscoelasticity, no toxic residue, and strong applicability.

[0084] Examples 9-13

[0085] Examples 9-13 describe the cross-linking reaction preparation of sodium hyaluronate of varying molecular weights and concentrations with two-arm polyethylene glycol (2arm-NH2-PEG-COOH), and the elastic modulus of the gels after 24 hours of reaction. Unlike Examples 1-8, the NH2-PEG-COOH was capped with different functional groups (amino and carboxyl). During the cross-linking reaction, the amino and carboxyl groups, respectively, coupled with the carboxyl and hydroxyl groups on the sodium hyaluronate, ultimately yielding the corresponding dual-cross-linked polymers.

[0086] 1. Sodium hyaluronate of different concentrations and molecular weights was dissolved in 40 mL of phosphate buffer solution with 0.03 g of 2arm-NH2-PEG-COOH (molecular weight: 300 Da) and 0.1 g of DMTMM. The concentrations and weight-average molecular weights of sodium hyaluronate were 1% and 2 million Da (Example 9), 2% and 2 million Da (Example 10), 3% and 2 million Da (Example 11), 2% and 1.35 million Da (Example 12), and 2% and 2.8 million Da (Example 13).

[0087] 2. Use a homogenizer to homogenize for 2 hours to dissolve and homogenize the raw materials in step 1.

[0088] 3. The homogeneous solution obtained in step 2 was placed in an oven at 25°C for crosslinking for 24 hours to obtain a solid gel, designated as sample 9-13. The structure of sample 9-13 can be referred to as formula II, and the degree of polymerization x of the polymer ranges from 2000 to 5000.

[0089] 4. Rheological properties of samples 9, 10, 11, 12, and 13 obtained in step 3 were tested for elastic modulus. The results are shown in Table 3. Table 3 shows that, compared with the test results of Examples 9-11, the elastic modulus of the gel significantly increased with increasing sodium hyaluronate concentration, with no crosslinking reaction occurring with 1% sodium hyaluronate and the crosslinking agent. Compared with the test results of Examples 10, 12, and 13, the elastic modulus of the gel increased with increasing sodium hyaluronate molecular weight, and the degree of crosslinking also increased.

[0090] Table 3 Elastic modulus of gels cross-linked with sodium hyaluronate and NH2-PEG-COOH at different concentrations and molecular weights

[0091] The above describes the embodiments of the present invention. However, the present invention is not limited to the above embodiments. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present invention shall be included in the scope of protection of the present invention.

Claims

1. A polymer, characterized in that The polymer is formed by cross-linking hyaluronic acid or a hyaluronic acid derivative with a PEG derivative, and the polymer has a structure as shown in Formula I: A is provided by a PEG derivative, wherein the PEG derivative is selected from a multi-arm PEG terminated with a functional group, wherein the functional group is selected from at least one of a hydroxyl group, an amino group and a carboxyl group, and the multi-arm refers to no less than two arms; Y is a connecting group selected from a group with strong polarity, preferably at least one of an ester group, an amide group, etc.; G is provided by hyaluronic acid or its derivatives; 1≤m≤the number of functional groups of the PEG derivative.

2. The polymer according to claim 1, characterized in that The multi-arm is selected from at least one of two arms to sixteen arms; Preferably, the PEG derivative is selected from at least one of two-arm PEG, three-arm PEG, four-arm PEG, five-arm PEG, six-arm PEG, seven-arm PEG, eight-arm PEG, nine-arm PEG, ten-arm PEG, eleven-arm PEG, twelve-arm PEG, thirteen-arm PEG, fourteen-arm PEG, fifteen-arm PEG, sixteen-arm PEG, etc. terminated with hydroxyl, amino and / or carboxyl groups; Preferably, the PEG derivative is selected from two-arm PEG, four-arm PEG or eight-arm PEG terminated with hydroxyl, amino and / or carboxyl groups; Also preferably, the PEG derivative is selected from hydroxyl-terminated eight-arm PEG, carboxyl-terminated eight-arm PEG, amino-terminated two-arm PEG, amino-terminated eight-arm PEG, and eight-arm PEG terminated with both hydroxyl and amino groups.

3. The polymer according to claim 1 or 2, characterized in that The weight average molecular weight of the hyaluronic acid or its derivative is 1 million Da to 5 million Da, preferably 1.5 million Da to 5 million Da; preferably, the weight average molecular weight of the hyaluronic acid or its derivative is 1.80 Da to 3 million Da; Preferably, m is an integer.

4. The polymer according to any one of claims 1 to 3, characterized in that The molecular weight of the PEG derivative is 100-100000Da; The hyaluronic acid derivative is sodium hyaluronate.

5. The polymer according to any one of claims 1 to 4, characterized in that The degree of polymerization x of the polymer is selected from 1000-8000.

6. The method for preparing the polymer according to any one of claims 1 to 5, characterized in that: The preparation method comprises the following steps: mixing PEG derivatives, hyaluronic acid or its derivatives, a condensing agent and a buffer, homogenizing the mixture to obtain a uniform solution, and performing a cross-linking reaction to obtain the polymer.

7. A gel, characterized in that: The gel comprises the polymer according to any one of claims 1 to 5 and a liquid, the polymer has a cross-linked network structure, and the liquid is stored in the network structure; preferably, the liquid is water, that is, the gel is a hydrogel.

8. A composition, characterized in that The composition comprises the polymer or the gel according to any one of claims 1 to 5.

9. A preparation, characterized in that The preparation contains the polymer, gel or composition according to any one of claims 1-5.

10. Use of the polymer according to any one of claims 1 to 5, the gel according to claim 7 or the composition according to claim 8 in the preparation of a product, wherein the product is any one of medicines, pharmaceutical excipients, medical devices, cosmetics, cosmetic raw materials, skin care products, skin care raw materials, women's care products, daily necessities, foods, food additives, dietary supplements, health products, meal replacements, dietary supplements, veterinary medicines, veterinary foods, veterinary medical devices, veterinary care products, etc., or is an industrial product.

11. Use of the polymer according to any one of claims 1 to 5, the gel according to claim 7 or the composition according to claim 8 in the preparation of any product for tissue regeneration, tissue repair, tissue filling, wound care, tissue adhesion prevention, postoperative care, wrinkle removal, plastic surgery, medical cosmetology, daily beauty, anti-aging, whitening and skin care, scalp care, hair transplantation, hair growth, fever reduction, lubrication, etc.

12. Use of the polymer according to any one of claims 1 to 5, the gel according to claim 7 or the composition according to claim 8 in the preparation of a medicament for preventing and / or treating diseases related to overweight; Preferably, the diseases associated with overweight include at least one of overweight, obesity, prediabetes, diabetes, hypercholesterolemia, hypertension, cardiovascular and cerebrovascular diseases, constipation, osteoarthritis, gout, sleep apnea syndrome, infertility, menstrual disorders, abnormal ovulation, abnormal hormone levels, etc.

13. A method for preventing and / or treating diseases associated with overweight, comprising administering a suitable dose of the polymer according to any one of claims 1 to 5, the gel according to claim 7, the composition according to claim 8, or the preparation according to claim 9 to a subject in need thereof; Preferably, the diseases associated with overweight include at least one of overweight, obesity, prediabetes, diabetes, hypercholesterolemia, hypertension, cardiovascular and cerebrovascular diseases, constipation, osteoarthritis, gout, sleep apnea syndrome, infertility, menstrual disorders, abnormal ovulation, abnormal hormone levels, etc.

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