Multifunctional light-responsive hydrogel, raw material thereof, preparation method and application thereof

Abstract

The present application relates to a kind of multifunctional photoresponsive hydrogel, its raw material, preparation method and application, by introducing carbon-carbon double bond and special functional group on natural polymer skeleton, obtain natural polymer derivative, and the natural polymer derivative, crosslinking agent containing multiple mercapto and photo initiator are combined, natural polymer derivative, crosslinking agent can be under the action of photo initiator, after light, crosslinking reaction occurs and forms multifunctional photoresponsive hydrogel.The present application is improved by structure design and functionalization to photoresponsive hydrogel, and endows hydrogel with more powerful anti-ultraviolet, anti-inflammatory, antibacterial, antioxidant, repair promotion and other effects.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of skin care products, and particularly relates to a multifunctional light-responsive hydrogel, raw materials, a preparation method and application thereof. BACKGROUND

[0002] A hydrogel is a soft material composed of a hydrophilic polymer network, which can absorb and retain a large amount of water while maintaining its three-dimensional structure. Due to its unique biocompatibility, controllable physicochemical properties and biomimetic extracellular matrix characteristics, hydrogels have wide applications in biomedical, tissue engineering, drug delivery, flexible electronics and other fields. In recent years, hydrogels have developed from static materials to "intelligent" materials that adapt to various stimuli (such as pH, temperature, chemicals, electricity or light). Among them, light-responsive hydrogels have become a research hotspot in intelligent materials due to their high spatiotemporal precision, non-invasive control and programmability. However, pure light-responsive hydrogels have relatively single functions, which are difficult to meet the needs of actual multi-application. The existing light-responsive hydrogels lack structural design and functional modification. In addition to moisturizing and isolation, hydrogels are difficult to meet the special needs of other applications, such as anti-ultraviolet, promoting tissue regeneration and repair, and antioxidant. SUMMARY

[0003] In view of the technical problems existing in the prior art, the application provides a multifunctional light-responsive hydrogel, raw materials, a preparation method and application thereof. The light-responsive hydrogel is structurally designed and functionally improved, so as to endow the hydrogel with stronger anti-ultraviolet, anti-inflammatory, antibacterial and antioxidant effects.

[0004] In a first aspect of the application, a natural polymer derivative is provided, which has a structure as shown in Formula I:

[0005] In Formula I, is a natural polymer skeleton, P 1、 P2 is independently selected from a dehydroxyl residue, a decarboxyl residue and a deaminated residue of a natural polymer repeating unit or fragment, R1 is a group containing a carbon-carbon double bond; R2 is any one of an ultraviolet absorption small molecule, a small molecule containing a polyphenol structure or a short peptide with a repair-promoting function, X and Y are independently selected from an ether bond, an ester bond, an amide bond and a coupling molecule, the coupling molecule includes at least two grafting sites of amino and / or hydrazine, is connected to a carboxyl site on the natural polymer through an amidation reaction at one end, and is connected to R1 or R2 at the other end; m and n are independently selected from integers of 1-10000;

[0006] The natural polymer includes at least one of sodium hyaluronate, sodium alginate, chitosan, gelatin, collagen, chondroitin sulfate or fibrin; ​

[0007] Preferably, the natural polymer has a molecular weight of 2 kDa to 5000 kDa.

[0008] In some embodiments of the present application, the R1 modification rate of the natural polymer is 10% to 50%.

[0009] In some embodiments of the present application, the R2 modification rate of the natural polymer is 5% to 50%.

[0010] In a second aspect of the present application, a multifunctional light-responsive hydrogel is provided, which is prepared from component A, component B and a photoinitiator in a solvent, wherein component A is the natural polymer derivative described above, component B is a crosslinking agent containing multiple thiol groups, and the photoinitiator is a small molecule UV absorber.

[0011] Preferably, the concentration of component A is 1 to 5 wt%, the concentration of component B is 1 to 5 wt%, the concentration of the photoinitiator is 0.001 to 1 wt%, and the rest is the solvent.

[0012] Preferably, the crosslinking agent containing multiple thiol groups is selected from dithiothreitol, thiolated polyethylene glycol, and thiol-modified biological macromolecules.

[0013] Further preferably, the biological macromolecule is any one of hyaluronic acid, gelatin, collagen, alginic acid, and chitosan.

[0014] Preferably, the solvent is selected from distilled water, physiological saline, or a buffer solution.

[0015] In some embodiments of the present application, the group containing a carbon-carbon double bond is selected from a vinyl group, a propenyl group, an allyloxy group, a cyclohexenyl group, a norbornenyl group, and a bicyclo[2.2.2]oct-5-enyl group.

[0016] In some embodiments of the present application, the small molecule UV absorber is a carboxyl-containing small molecule sunscreen.

[0017] Preferably, the carboxyl-containing small molecule sunscreen includes cinnamic acid or a cinnamic acid derivative; the cinnamic acid derivative can undergo photodimerization to form a crosslinked network under irradiation of a specific wavelength, thereby improving the mechanical properties of the light-responsive hydrogel.

[0018] Preferably, the carboxyl-containing small molecule sunscreen includes at least one of salicylic acid, a salicylic acid derivative, benzoic acid, a benzoic acid derivative, 2,6-naphthalene dicarboxylic acid, or 1,8-dihydroxy-3-carboxy anthraquinone. These small molecules with strong UV absorption endow the light-responsive hydrogel with the effects of anti-UV, anti-inflammatory, and anti-oxidation.

[0019] In some embodiments of the present application, the small molecule containing polyphenol structure is selected from any one of dopamine, dopamine derivative, 3,4-dihydroxybenzoic acid, 3,4-dihydroxybenzoic acid derivative, gallic acid, gallic acid derivative. These small molecules containing polyphenol structure can endow the photoresponsive hydrogel with antibacterial, antioxidant, wet tissue adhesion and other functions.

[0020] In some embodiments of the present application, the short peptide with repair-promoting function includes at least one of EGF short peptide, oligopeptide-1, QK peptide, PR1P peptide, RGD peptide, BDNF mimic peptide or BMP-2 derivative peptide. These short peptides with repair-promoting function can endow the photoresponsive hydrogel with functions of promoting tissue regeneration and repair.

[0021] In some embodiments of the present application, the photoinitiator includes at least one of lithium phenyl(2,4,6-trimethylbenzoyl)phosphinate (LAP), 2-hydroxy-2-methyl-1-[4-(2-hydroxyethoxy)phenyl]-1-propanone (I2959), eosin Y, α-ketoglutaric acid, 2,4,6-trimethylbenzoyl diphenyl phosphine oxide or 1-hydroxycyclohexyl phenyl ketone.

[0022] In a third aspect of the present application, a preparation method of the natural polymer derivative with the structure shown in formula I is provided, and any one of the following preparation methods a-d is selected:

[0023] a. R1 and R2 are sequentially modified at the hydroxyl, carboxyl or amino site of the natural polymer;

[0024] b. an amino group and / or a hydrazide-containing coupling molecule is introduced at the carboxyl site of the natural polymer, and then R1 and R2 are sequentially modified at the amino group and / or the hydrazide;

[0025] c. R1 is modified at the carboxyl or hydroxyl site of the natural polymer, an amino group-containing coupling agent is introduced at the carboxyl site of the natural polymer derivative modified by R1, and then R2 is modified at the amino group;

[0026] d. R2 is modified at the carboxyl or hydroxyl site of the natural polymer, an amino group and / or hydrazide-containing coupling agent is introduced at the carboxyl site of the natural polymer derivative modified by R2, and then R1 is modified at the amino group or hydrazide;

[0027] Preferably, the coupling molecule is selected from any one of ethylenediamine, propylenediamine, butylenediamine, pentanediamine, hexanediamine, heptanediamine, octanediamine, nonanediamine, decanediamine, amino-PEG, oxalic acid dihydrazide, malonic acid dihydrazide, succinic acid dihydrazide, glutaric acid dihydrazide, adipic acid dihydrazide, pimelic acid dihydrazide, suberic acid dihydrazide, azelaic acid dihydrazide and sebacic acid dihydrazide.

[0028] In a fourth aspect, the present application provides a method for preparing the multifunctional photoresponsive hydrogel, comprising the following steps:

[0029] The component A, component B and photoinitiator are dissolved in a solvent to obtain a hydrogel precursor solution:

[0030] The hydrogel precursor solution is irradiated by a light source to form a hydrogel through photocrosslinking.

[0031] In a fifth aspect, the present application provides the application of the multifunctional photoresponsive hydrogel, which is selected from the following applications:

[0032] The photoresponsive hydrogel is applied in a product for promoting tissue regeneration and repair;

[0033] The photoresponsive hydrogel is applied in an ultraviolet protection product;

[0034] The photoresponsive hydrogel is applied in an antioxidant product;

[0035] The photoresponsive hydrogel is applied in a disinfectant product.

[0036] The multifunctional photoresponsive hydrogel, raw material, preparation method and application disclosed by the embodiments of the present application are based on a natural polymer derivative with a structure shown in formula I, which has a carbon-carbon double bond and a special functional group on the skeleton. The photoresponsive hydrogel comprises a natural polymer derivative with a structure shown in formula I (component A), a crosslinking agent containing multiple mercapto groups (component B) and a photoinitiator. The two components A and B can be crosslinked to form a hydrogel under the action of the photoinitiator after irradiation. At this time, the functional groups on the natural polymer derivative with a structure shown in formula I can enhance the function at the application site, and the crosslinking network can increase the residence time of the functional groups and prolong the period of the functional groups to play the efficacy. For example, when the functional group is cinnamic acid or a cinnamic acid derivative; the cinnamic acid derivative will undergo photodimerization to form a crosslinking network under irradiation of a specific wavelength, which can improve the mechanical properties of the photoresponsive hydrogel, thereby enhancing the waterproof performance.

[0037] In summary, by introducing different functional groups into the molecular skeleton of the photoresponsive hydrogel, the photoresponsive intelligent hydrogel with different efficacies is designed, and the performance of the photoresponsive hydrogel in terms of ultraviolet resistance, anti-inflammatory, antibacterial, antioxidant, enhanced tissue adhesion, promotion of tissue repair and regeneration, etc. can be improved to flexibly meet the needs of different application scenarios. The present application can introduce functional groups into the molecular skeleton of the hydrogel through a general method by specially designing the structure of the two-component photoresponsive hydrogel, so as to endow the photoresponsive hydrogel with more specific functions, such as stronger ultraviolet resistance, anti-inflammatory, antibacterial, antioxidant, repair promotion, etc., to meet the individualized needs of different application fields.

[0038] It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the present disclosure, as claimed. BRIEF DESCRIPTION OF DRAWINGS

[0039] The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate embodiments consistent with the present application and, together with the description, serve to explain the principles of the application;

[0040] Figure 1 Figure 1 is a test chart of the tissue adhesion strength of a small molecule modified photoresponsive hydrogel containing a polyphenol structure according to an embodiment of the present application;

[0041] Figure 2 Figure 2 is a test chart of the burst strength of a small molecule modified photoresponsive hydrogel containing a polyphenol structure according to an embodiment of the present application;

[0042] Figure 3 Figure 3 is a comparison chart of the effect of a short peptide modified photoresponsive hydrogel with a repair promoting function on promoting skin repair according to an embodiment of the present application. DETAILED DESCRIPTION

[0043] The concept and technical effects of the present application will be described below in conjunction with the embodiments to make the purpose, features and effects of the present application clear. Obviously, the described embodiments are only some of the embodiments of the present application, but not all the embodiments. Based on the embodiments of the present application, other embodiments obtained by those skilled in the art without creative labor are within the scope of protection of the present application.

[0044] The experimental methods not specified in the embodiments are usually carried out according to the conventional conditions in the art or according to the recommended conditions of the manufacturers; the raw materials, reagents, etc. used, if not specifically stated, are all raw materials and reagents that can be obtained through commercial channels.

[0045] Example 1: Preparation of sodium hyaluronate derivative A1

[0046] (1) 2.0 g of sodium hyaluronate (MW ≈ 400 kDa) was dissolved in 400 mL of MES solution, the pH was adjusted to 4-5, 8.37 g of adipic acid dihydrazide and 1.05 g of EDC were added, the pH was maintained in the range of 4-5, and the stirring was continued overnight. After the reaction was completed, the hydrazide functionalized sodium hyaluronate was dialyzed in a dialysis bag (MWCO = 10 kDa) in deionized water for three days, and freeze-dried to obtain the hydrazide functionalized sodium hyaluronate.

[0047] (2) 5 g of p-nitrocinnamic acid was dissolved in 50 mL of N, N'-dimethylformamide (DMF) at 40 °C, and then equimolar amounts of NHS and EDC were added. The reaction was carried out in the dark for 24 h. After the reaction was completed, the by-product dicyclohexylurea was removed by filtration, and the filtrate was precipitated in water to obtain a solid. The solid was vacuum dried to obtain N-succinimidyl p-nitrocinnamate with a yield of about 90%.

[0048] (3) 1.0 g of hydrazide-functionalized sodium hyaluronate was dissolved in 200 mL of water, and the pH was adjusted to 7.0. Then, 4.33 g of norbornene diacid anhydride and 2.00 g of N-succinimidyl p-nitrocinnamate were added, and the reaction was stirred overnight. Subsequently, dialysis was performed in deionized water using a dialysis bag (MWCO = 10 kDa) for three days, and freeze-drying was performed to obtain a norbornene and p-nitrocinnamic acid-modified sodium hyaluronate derivative with a yield of about 74%. Nuclear magnetic resonance detection showed that the norbornene modification rate was about 24%, and the p-nitrocinnamic acid modification rate was 11%.

[0049] Example 2: Preparation of sodium alginate derivative A2;

[0050] (1) 2.0 g of sodium alginate (MW ≈ 200 kDa) was dissolved in 40 mL of 0.1 M NaOH solution, and 1.7 g of allyl glycidyl ether was added. The reaction was continuously stirred at 35 °C for 24 h. After the reaction was completed, the reaction solution was poured into 250 mL of ice ethanol to precipitate, and the precipitate was filtered. After being redissolved in water, dialysis was performed in deionized water using a dialysis bag (MWCO = 10 kDa) for three days, and freeze-drying was performed to obtain allyl-functionalized sodium alginate.

[0051] (2) 2.0 g of allyl-functionalized sodium alginate was dissolved in 200 mL of MES solution, and 4.65 g of butanediamine and 2.0 g of EDC were added. The pH was adjusted to 4.75, and the stirring was continuously performed overnight. After the reaction was completed, dialysis was performed in deionized water using a dialysis bag (MWCO = 10 kDa) for three days, and freeze-drying was performed to obtain amino and allyl-functionalized sodium alginate.

[0052] (3) 5 g of p-methoxycinnamic acid was dissolved in 50 mL of N, N'-dimethylformamide (DMF) at 40 °C, and then equimolar amounts of NHS and EDC were added. The reaction was carried out in the dark for 24 h. After the reaction was completed, the by-product dicyclohexylurea was removed by filtration, and the filtrate was precipitated in water to obtain a solid. The solid was vacuum dried to obtain N-succinimidyl p-nitrocinnamate with a yield of about 90%.

[0053] (4) Take 1.0 g of amino and allyl functionalized sodium alginate dissolved in 200 mL of water, adjust the pH to 7.0, add 2.00 g of N-succinimidyl p-methoxycinnamate, stir overnight, then dialyze in a dialysis bag (MWCO = 10 kDa) in deionized water for three days, freeze-dry to obtain an allyl and p-methoxycinnamic acid modified sodium alginate derivative, with a yield of about 71%, and a nuclear magnetic resonance detection of about 33% allyl modification rate and 9% p-methoxycinnamic acid modification rate.

[0054] Example 3: Preparation of sodium hyaluronate derivative A3;

[0055] (1) Take 2.0 g of sodium hyaluronate (MW ≈ 100 kDa) dissolved in 200 mL of MES buffer solution, adjust the pH to 5.5, add 4.4 g of 4-(4,6-dimethoxytriazin-2-yl)-4-methyl morpholine hydrochloride and 1.23 g of norbornene-2-methylamine, continuously stir at 20°C for 24 h, after the reaction is completed, pour the reaction solution into 500 mL of ice ethanol to precipitate, filter out the precipitate, redissolve with water, then dialyze in a dialysis bag (MWCO = 10 kDa) in deionized water for three days, freeze-dry to obtain norbornene functionalized sodium hyaluronate.

[0056] (2) Take 2.0 g of norbornene functionalized sodium hyaluronate dissolved in 200 mL of MES solution, add 18.37 g of adipic acid dihydrazide and 4.05 g of EDC, adjust the pH to 4.75, continuously stir overnight, after the reaction is completed, dialyze in a dialysis bag (MWCO = 10 kDa) in deionized water for three days, freeze-dry to obtain hydrazide and norbornene functionalized sodium hyaluronate.

[0057] (3) Dissolve 5 g of cinnamic acid in 50 mL of N, N'-dimethylformamide (DMF) at 40°C, then add equimolar amounts of NHS and EDC, avoid light reaction for 24 h, after the reaction is completed, filter out the generated by-product dicyclohexylurea, pour the filtrate into water to precipitate, vacuum dry the obtained solid to obtain N-succinimidyl cinnamate, with a yield of about 91%.

[0058] (4) Take 1.0 g of hydrazide and norbornene functionalized sodium hyaluronate dissolved in 200 mL of water, adjust the pH to 7.0, add 2.00 g of N-succinimidyl cinnamate, stir overnight, then dialyze in a dialysis bag (MWCO = 10 kDa) in deionized water for three days, freeze-dry to obtain norbornene and cinnamic acid modified sodium hyaluronate derivative, with a yield of about 82%, and a nuclear magnetic resonance detection of about 27% norbornene modification rate and 12% cinnamic acid modification rate.

[0059] Example 4: Preparation of gelatin derivative A4

[0060] Take 10.0 g of gelatin (MW ≈ 500 kDa) and dissolve it in 200 mL of 0.5 M NaHCO3 solution at 50°C, adjust the pH to 8-9, add 10 g of norbornene anhydride, continue stirring overnight, after the reaction is completed, use dialysis bag (MWCO = 10 kDa) to dialyze in deionized water for three days, freeze-drying to obtain norbornene functionalized gelatin.

[0061] (2) At 40°C, 5 g of o-hydroxybenzoic acid was dissolved in 50 mL of N, N'-dimethylformamide (DMF), and then an equimolar amount of NHS and EDC was added, and the reaction was carried out in the dark for 24 h. After the reaction was completed, the generated by-product dicyclohexylurea was removed by filtration, and the filtrate was poured into water to precipitate. The obtained solid was vacuum dried and recrystallized with DMF / ethanol to obtain N-succinimidyl o-hydroxybenzoate.

[0062] (3) Take 5.0 g of norbornene functionalized gelatin and dissolve it in 100 mL of water at 50°C, adjust the pH to 7.0, add 2.00 g of N-succinimidyl o-hydroxybenzoate, stir and react overnight, then dialyze in deionized water with dialysis bag (MWCO = 10 kDa) for three days, freeze-drying to obtain norbornene and o-hydroxybenzoic acid modified gelatin derivative, with a yield of about 78%, and the norbornene modification rate detected by nuclear magnetic resonance is about 31%, and the o-hydroxybenzoic acid modification rate is 16%.

[0063] Example 5 Preparation of collagen derivative A5

[0064] (1) Take 2.0 g of bovine collagen (MW ≈ 300 kDa) and dissolve it in 50 mL of 0.5 M NaHCO3 solution at 50°C, adjust the pH to 8-9, add 10 g of norbornene anhydride, continue stirring overnight, after the reaction is completed, use dialysis bag (MWCO = 10 kDa) to dialyze in deionized water for three days, freeze-drying to obtain norbornene functionalized collagen.

[0065] (2) At 40°C, 5 g of 2,6-naphthalene dicarboxylic acid was dissolved in 50 mL of N, N'-dimethylformamide (DMF), and then an equimolar amount of NHS and EDC was added, and the reaction was carried out in the dark for 24 h. After the reaction was completed, the generated by-product dicyclohexylurea was removed by filtration, and the filtrate was poured into water to precipitate. The obtained solid was vacuum dried and recrystallized with DMF / ethanol to obtain N-succinimidyl 2,6-naphthalene dicarboxylate.

[0066] (3) Take 1.0 g of norbornene functionalized gelatin, dissolve in 50 mL of water at 50°C, adjust the pH to 7.0, add 2.00 g of N-succinimidyl 2,6-naphthalene dicarboxylate, stir overnight, then dialyze in a dialysis bag (MWCO = 10 kDa) in deionized water for three days, freeze-dry to obtain a norbornene and 2,6-naphthalene dicarboxylic acid modified collagen derivative, with a yield of about 72%, and a norbornene modification rate of about 24% and a 2,6-naphthalene dicarboxylic acid modification rate of 11% detected by nuclear magnetic resonance.

[0067] Example 6 Preparation of chitosan derivative A6

[0068] (1) Take 2.0 g of carboxymethyl chitosan (MW ≈ 200 kDa) and dissolve in 100 mL of 0.5 M NaHCO3 solution, adjust the pH to 8-9, add 10 g of norbornene dianhydride, continue stirring overnight, after the reaction is completed, dialyze in a dialysis bag (MWCO = 10 kDa) in deionized water for three days, freeze-dry to obtain norbornene functionalized chitosan.

[0069] (2) Dissolve 5 g of 1,8-dihydroxy-3-carboxy anthraquinone in 50 mL of N, N'-dimethylformamide (DMF) at 40°C, then add equimolar amounts of NHS and EDC, avoid light reaction for 24 h, after the reaction is completed, filter to remove the generated by-product dicyclohexyl urea, pour the filtrate into water to precipitate, vacuum dry the obtained solid, then recrystallize with DMF / ethanol to obtain N-succinimidyl 1,8-dihydroxy-3-carboxy anthraquinone ester.

[0070] (3) Take 1.0 g of norbornene functionalized chitosan and dissolve in 100 mL of deionized water, adjust the pH to 7.0, add 2.00 g of N-succinimidyl 1,8-dihydroxy-3-carboxy anthraquinone ester, stir overnight, then dialyze in a dialysis bag (MWCO = 10 kDa) in deionized water for three days, freeze-dry to obtain a norbornene and 1,8-dihydroxy-3-carboxy anthraquinone modified chitosan derivative, with a yield of about 67%, and a norbornene modification rate of about 30% and a 1,8-dihydroxy-3-carboxy anthraquinone modification rate of 7% detected by nuclear magnetic resonance.

[0071] Example 7 Preparation of chondroitin sulfate derivative A7

[0072] (1) 2.0 g of chondroitin sulfate was weighed and dissolved in 200 mL of MES buffer solution, the pH was adjusted to 5.5, 4.4 g of 4-(4,6-dimethoxytriazin-2-yl)-4-methyl morpholine hydrochloride and 1.5 g of 3-cyclohexenyl-1-amine were added, and stirring was continued at 20°C for 24 h. After the reaction was completed, the reaction solution was poured into 500 mL of ice ethanol for precipitation, and the precipitate was filtered. After re-dissolving with water, dialysis was performed in deionized water for three days using a dialysis bag (MWCO = 10 kDa), and freeze-drying was performed to obtain 3-cyclohexenyl-functionalized chondroitin sulfate.

[0073] (2) 0.5 g of 3-cyclohexenyl-functionalized chondroitin sulfate was weighed and dissolved in 150 mL of MES solution, 4.24 g of pentanediamine and 2.05 g of EDC were added, the pH was adjusted to 4.75, and stirring was continued overnight. After the reaction was completed, dialysis was performed in deionized water for three days using a dialysis bag (MWCO = 10 kDa), and freeze-drying was performed to obtain amino and 3-cyclohexenyl-functionalized chondroitin sulfate.

[0074] (3) 5 g of p-methoxysalicylic acid was dissolved in 50 mL of N, N'-dimethylformamide (DMF) at 40°C, and an equimolar amount of NHS and EDC was added, and the reaction was carried out in the dark for 24 h. After the reaction was completed, the generated by-product dicyclohexyl urea was filtered out, the filtrate was poured into water for precipitation, and the obtained solid was vacuum dried and recrystallized with DMF / ethanol to obtain N-succinimidyl p-methoxysalicylate.

[0075] (4) 1.0 g of amino and 3-cyclohexenyl-functionalized chondroitin sulfate was weighed and dissolved in 100 mL of water, the pH was adjusted to 7.0, 1.8 g of N-succinimidyl p-methoxysalicylate was added, and stirring was continued overnight. Then, dialysis was performed in deionized water for three days using a dialysis bag (MWCO = 10 kDa), and freeze-drying was performed to obtain 3-cyclohexenyl and p-methoxysalicylic acid-modified chondroitin sulfate derivatives, with a yield of about 78%, and 3-cyclohexenyl modification rate of about 22% and salicylic acid modification rate of 10% detected by nuclear magnetic resonance.

[0076] Example 8: Preparation of sodium hyaluronate derivative A8;

[0077] (1) Take 2.0 g of sodium hyaluronate, dissolve it in 200 mL of MES buffer solution, adjust the pH to 5.5, add 4.4 g of 4-(4,6-dimethoxytriazin-2-yl)-4-methylmorpholine hydrochloride and 1.9 g of bicyclo[2.2.2]oct-5-ene-2-methylamine, continuously stir at 20°C for 24 h, after the reaction is completed, pour the reaction solution into 500 mL of ice ethanol to precipitate, filter out the precipitate, redissolve it with water, then use a dialysis bag (MWCO = 10 kDa) to dialyze in deionized water for three days, freeze-dry to obtain bicyclo[2.2.2]oct-5-ene functionalized sodium hyaluronate.

[0078] (2) Take 1.5 g of bicyclo[2.2.2]oct-5-ene functionalized sodium hyaluronate, dissolve it in 150 mL of MES solution, add 4.24 g of hexanediamine and 2.05 g of EDC, adjust the pH to 4.75, continuously stir overnight, after the reaction is completed, use a dialysis bag (MWCO = 10 kDa) to dialyze in deionized water for three days, freeze-dry to obtain amino and bicyclo[2.2.2]oct-5-ene functionalized sodium hyaluronate.

[0079] (3) Dissolve 5 g of salicylic acid in 50 mL of N, N'-dimethylformamide (DMF) at 40°C, then add equimolar amounts of NHS and EDC, avoid light reaction for 24 h, after the reaction is completed, filter out the generated by-product dicyclohexylurea, pour the filtrate into water to precipitate, vacuum dry the obtained solid, then recrystallize it with DMF / ethanol to obtain N-succinimidyl salicylate.

[0080] (4) Take 1.0 g of amino and bicyclo[2.2.2]oct-5-ene functionalized sodium hyaluronate, dissolve it in 100 mL of water, adjust the pH to 7.0, add 1.8 g of N-succinimidyl salicylate, stir and react overnight, then use a dialysis bag (MWCO = 10 kDa) to dialyze in deionized water for three days, freeze-dry to obtain bicyclo[2.2.2]oct-5-ene and salicylic acid modified sodium hyaluronate derivative, the yield is about 75%, the 3-cyclohexene modification rate is about 27% and the salicylic acid modification rate is 13% as detected by nuclear magnetic resonance.

[0081] Example 9: Preparation of sodium hyaluronate derivative A9;

[0082] (1) Take 2.0 g of sodium hyaluronate (MW ≈ 800 kDa), dissolve it in 40 mL of 0.1M NaOH solution, add 1.7 g of allyl glycidyl ether, continuously stir at 35°C for 24 h, after the reaction is completed, pour the reaction solution into 250 mL of ice ethanol to precipitate, filter out the precipitate, redissolve it with water, then use a dialysis bag (MWCO = 10 kDa) to dialyze in deionized water for three days, freeze-dry to obtain allyl functionalized sodium hyaluronate.

[0083] (2) 2.0 g of allyl-functionalized sodium hyaluronate was weighed and dissolved in 200 mL of MES solution, 10.00 g of double-end amino-terminated polyethylene glycol (H2N-PEG 1000 -NH2) and 2.0 g of EDC were added, the pH was adjusted to 4.75, and stirring was continued overnight. After the reaction was completed, dialysis bags (MWCO = 10 kDa) were used for dialysis in deionized water for three days, and freeze-drying was performed to obtain amino-PEG and allyl-functionalized sodium hyaluronate.

[0084] (3) 1.0 g of amino- and allyl-functionalized sodium hyaluronate was weighed and dissolved in 200 mL of water, the pH was adjusted to 7.0, 2.00 g of N-succinimidyl p-methoxycinnamate was added, and stirring was continued overnight. Then dialysis bags (MWCO = 10 kDa) were used for dialysis in deionized water for three days, and freeze-drying was performed to obtain an allyl- and p-methoxycinnamate-modified sodium hyaluronate derivative, with a yield of about 71%, an allyl modification rate of about 33% detected by nuclear magnetic resonance, and a p-methoxycinnamate modification rate of 9%.

[0085] Example 10: Preparation of hyaluronic acid derivative A10;

[0086] (1) 2.0 g of sodium hyaluronate (MW ≈ 50 kDa) was weighed and dissolved in 200 mL of a MES buffer solution, the pH was adjusted to 5.5, 4.4 g of 4-(4,6-dimethoxytriazin-2-yl)-4-methylmorpholinium hydrochloride and 1.23 g of norbornene-2-methylamine were added, and stirring was continued at 20°C for 24 h. After the reaction was completed, the reaction solution was poured into 500 mL of ice ethanol to precipitate, the precipitate was filtered, redissolved in water, dialysis bags (MWCO = 10 kDa) were used for dialysis in deionized water for three days, and freeze-drying was performed to obtain norbornene-functionalized sodium hyaluronate.

[0087] (2) 2.0 g of norbornene-functionalized sodium hyaluronate was weighed and dissolved in 200 mL of a MES buffer solution, the pH was adjusted to 5.5, 4.4 g of 4-(4,6-dimethoxytriazin-2-yl)-4-methylmorpholinium hydrochloride and 1.53 g of dopamine were added, and stirring was continued at 20°C for 24 h. After the reaction was completed, the reaction solution was poured into 500 mL of ice ethanol to precipitate, the precipitate was filtered, redissolved in water, dialysis bags (MWCO = 10 kDa) were used for dialysis in deionized water for three days, and freeze-drying was performed to obtain norbornene- and dopamine-functionalized sodium hyaluronate, with a yield of 69%, a norbornene modification rate of 27% detected by nuclear magnetic resonance, and a dopamine modification rate of 24%.

[0088] Example 11: Preparation of hyaluronic acid derivative A11;

[0089] (1) Take 2.0 g of sodium hyaluronate (MW ≈ 200 kDa) and dissolve it in 100 mL of 0.5 M NaHCO3 solution, adjust the pH to 8-9, add 10 g of norbornene anhydride, continue stirring overnight, after the reaction is completed, dialyze in a dialysis bag (MWCO = 10 kDa) in deionized water for three days, freeze-dry to obtain norbornene functionalized sodium hyaluronate.

[0090] (2) Take 2.0 g of norbornene functionalized sodium hyaluronate, dissolve it in 200 mL of MES buffer solution, adjust the pH to 5.5, add 4.4 g of 4-(4,6-dimethoxytriazin-2-yl)-4-methyl morpholine hydrochloride and 1.53 g of dopamine, continue stirring at 20°C for 24 h, after the reaction is completed, pour the reaction solution into 500 mL of ice ethanol to precipitate, filter out the precipitate, redissolve with water, dialyze in a dialysis bag (MWCO = 10 kDa) in deionized water for three days, freeze-dry to obtain norbornene and dopamine functionalized sodium hyaluronate, the yield is 73%, the norbornene modification rate is 28% and the dopamine modification rate is 17% measured by nuclear magnetic resonance.

[0091] Example 12: Preparation of alginate derivative A12;

[0092] (1) Take 2.0 g of sodium hyaluronate (MW ≈ 200 kDa) and dissolve it in 100 mL of 0.5 M NaHCO3 solution, adjust the pH to 8-9, add 10 g of norbornene anhydride, continue stirring overnight, after the reaction is completed, dialyze in a dialysis bag (MWCO = 10 kDa) in deionized water for three days, freeze-dry to obtain norbornene functionalized sodium hyaluronate.

[0093] (2) Dissolve 5 g of 3,4-dihydroxybenzoic acid in 50 mL of N, N'-dimethylformamide (DMF) at 40°C, then add equimolar amounts of NHS and EDC, avoid light reaction for 24 h, after the reaction is completed, filter out the generated by-product dicyclohexyl urea, pour the filtrate into water to precipitate, vacuum dry the obtained solid and recrystallize with DMF / ethanol to obtain N-succinimidyl 3,4-dihydroxybenzoate.

[0094] (3) Take 1.0 g of amino-functionalized alginate sodium and dissolve it in 200 mL of water, adjust the pH to 7.0, then add 4.33 g of norbornene anhydride and 2.00 g of N-succinimidyl 3,4-dihydroxybenzoate, stir and react overnight, then dialyze in a dialysis bag (MWCO = 10 kDa) in deionized water for three days, freeze-dry to obtain norbornene and 3,4-dihydroxybenzoic acid modified alginate sodium derivative, the yield is about 76%, the norbornene modification rate is 25% and the 3,4-dihydroxybenzoic acid modification rate is 13% measured by nuclear magnetic resonance.

[0095] Example 13: Preparation of hyaluronic acid derivative A13;

[0096] (1) Weigh 2.0 g of sodium hyaluronate (MW ≈ 200 kDa) and dissolve it in 40 mL of 0.1 M NaOH solution, add 1.7 g of allyl glycidyl ether, and continuously stir at 35°C for 24 h. After the reaction is completed, pour the reaction solution into 250 mL of ice ethanol to precipitate, filter out the precipitate, redissolve it with water, and dialyze it in deionized water for three days using a dialysis bag (MWCO = 10 kDa). Freeze-drying gives sodium hyaluronate functionalized with allyl groups.

[0097] (2) Weigh 2.0 g of sodium hyaluronate functionalized with allyl groups and dissolve it in 200 mL of MES solution, add 12.37 g of adipic acid dihydrazide and 2.0 g of EDC, adjust the pH to 4.75, and continuously stir overnight. After the reaction is completed, dialyze it in deionized water for three days using a dialysis bag (MWCO = 10 kDa), and freeze-dry to obtain sodium hyaluronate functionalized with both hydrazide and allyl groups.

[0098] (3) Dissolve 5 g of gallic acid in 50 mL of N, N'-dimethylformamide (DMF) at 40°C, and then add equimolar amounts of NHS and EDC, and react in the dark for 24 h. After the reaction is completed, filter out the generated byproduct dicyclohexylurea, pour the filtrate into water to precipitate, vacuum-dry the obtained solid, and recrystallize it from DMF / ethanol to obtain N-succinimidyl gallic acid ester.

[0099] (4) Weigh 1.0 g of sodium hyaluronate functionalized with both hydrazide and allyl groups and dissolve it in 200 mL of water, adjust the pH to 7.0, add 3.00 g of N-succinimidyl gallic acid ester, stir to react overnight, and then dialyze it in deionized water for three days using a dialysis bag (MWCO = 10 kDa). Freeze-drying gives sodium hyaluronate derivative functionalized with both allyl and gallic acid, with a yield of about 73%, an allyl modification rate of about 34% detected by NMR, and a gallic acid modification rate of 12%.

[0100] Example 14: Preparation of fibrin derivative A14;

[0101] (1) Weigh 10.0 g of fibrin (MW ≈ 500 kDa) and dissolve it in 200 mL of 0.2 M NaHCO3 solution at 50°C, adjust the pH to 8~9, add 10 g of norbornene diacid anhydride, and continuously stir overnight. After the reaction is completed, dialyze it in deionized water for three days using a dialysis bag (MWCO = 10 kDa), and freeze-dry to obtain norbornene-functionalized fibrin.

[0102] (2) 5 g of 3,4-dihydroxyphenylpropionic acid was dissolved in 50 mL of N, N'-dimethylformamide (DMF) at 40 °C, and then equimolar amounts of NHS and EDC were added, and the reaction was carried out in the dark for 24 h. After the reaction was completed, the by-product dicyclohexylurea generated was removed by filtration, and the filtrate was precipitated in water to obtain a solid. The obtained solid was vacuum dried and recrystallized with DMF / ethanol to obtain N-succinimidyl 3,4-dihydroxyphenylpropionate.

[0103] (3) 5.0 g of norbornene-functionalized fibrin was weighed and dissolved in 100 mL of water, and then 2.00 g of N-succinimidyl 3,4-dihydroxyphenylpropionate was added, and the reaction was stirred overnight. Then, dialysis bags (MWCO = 10 kDa) were used to dialyze in deionized water for three days, and then freeze-drying was performed to obtain a norbornene and 3,4-dihydroxyphenylpropionic acid modified fibrin derivative, with a yield of about 72%, a norbornene modification rate of 39%, and a 3,4-dihydroxyphenylpropionic acid modification rate of 21%.

[0104] Example 15: Preparation of gelatin derivative A15

[0105] (1) 10.0 g of gelatin (MW ≈ 500 kDa) was dissolved in 200 mL of a 0.2 M NaHCO3 solution at 50 °C, and the pH was adjusted to 8~9. Then, 10 g of norbornene diacid anhydride was added, and the reaction was continuously stirred overnight. After the reaction was completed, dialysis bags (MWCO = 10 kDa) were used to dialyze in deionized water for three days, and then freeze-drying was performed to obtain norbornene-functionalized gelatin.

[0106] (2) 5 g of 3,4-dihydroxyphenylacetic acid was dissolved in 50 mL of N, N'-dimethylformamide (DMF) at 40 °C, and then equimolar amounts of NHS and EDC were added, and the reaction was carried out in the dark for 24 h. After the reaction was completed, the by-product dicyclohexylurea generated was removed by filtration, and the filtrate was precipitated in water to obtain a solid. The obtained solid was vacuum dried and recrystallized with DMF / ethanol to obtain N-succinimidyl 3,4-dihydroxyphenylacetate.

[0107] (3) 5.0 g of norbornene-functionalized gelatin was weighed and dissolved in 100 mL of water, and then 2.00 g of N-succinimidyl 3,4-dihydroxyphenylacetate was added, and the reaction was stirred overnight. Then, dialysis bags (MWCO = 10 kDa) were used to dialyze in deionized water for three days, and then freeze-drying was performed to obtain a norbornene and 3,4-dihydroxyphenylacetate modified gelatin derivative, with a yield of about 71%, a norbornene modification rate of about 39%, and a 3,4-dihydroxyphenylacetate modification rate of 17%.

[0108] Example 16: Preparation of allyl-functionalized sodium hyaluronate a16 and hyaluronic acid derivative A16

[0109] (1) 2.0 g of sodium hyaluronate (MW = 200 kDa) was weighed and dissolved in 40 mL of 0.1 M NaOH solution, 2.0 g of allyl glycidyl ether was added, and stirring was continued at 35°C for 24 h. After the reaction was completed, the reaction solution was poured into 250 mL of ice ethanol to precipitate, and the precipitate was filtered. After re-dissolving with water, dialysis was performed in deionized water for three days using a dialysis bag (MWCO = 10 kDa), and freeze-drying was performed to obtain allyl-functionalized sodium hyaluronate a16.

[0110] (2) 1.0 g of allyl-functionalized sodium hyaluronate was weighed and dissolved in 200 mL of water, and the pH was adjusted to 7.0. 0.52 g of EDC and 0.52 g of N-hydroxysuccinimide were added, and stirring was continued for 30 min. After the reaction, EGF short peptide was added, and stirring was continued overnight. Then, dialysis was performed in deionized water for three days using a dialysis bag (MWCO = 10 kDa), and freeze-drying was performed to obtain an allyl and EGF short peptide-modified sodium hyaluronate derivative, with a yield of about 82%. The allyl modification rate was measured by NMR to be 33%, and the EGF short peptide modification rate was measured by UV quantification to be about 9%.

[0111] Examples 17-22: Preparation of hyaluronic acid derivatives A17-A22

[0112] The preparation methods of Examples 17-22 are different from Example 16 only in that equal amounts of oligopeptide-1, QK peptide, PR1P peptide, RGD peptide, BDNF mimetic peptide, and BMP-2 derivative peptide are sequentially used instead of EGF short peptide to replace EGF short peptide, to obtain sodium hyaluronate derivatives A17-A22.

[0113] The coupling molecules in the above examples are equivalent to any one selected from the group consisting of ethylenediamine, propylenediamine, butylenediamine, pentylenediamine, hexylenediamine, heptylenediamine, octylenediamine, nonylenediamine, decylenediamine, aminated PEG, oxalic acid dihydrazide, malonic acid dihydrazide, succinic acid dihydrazide, glutaric acid dihydrazide, adipic acid dihydrazide, pimelic acid dihydrazide, suberic acid dihydrazide, azelaic acid dihydrazide, and sebacic acid dihydrazide.

[0114] Example 23: Preparation of thiolated hyaluronic acid (HASH)

[0115] Take 2.0 g of hyaluronic acid sodium (MW ≈ 2000 kDa) and dissolve it in 200 mL of MES buffer, add 3.06 g of EDC and 0.96 g of N-hydroxysuccinimide, continue stirring for 30 min, then add 2.0 g of cysteamine hydrochloride, adjust the pH to 4.75, continue stirring for 24 h, after the reaction is completed, the reaction solution is dialyzed in a dialysis bag (MWCO = 10 kDa) in water containing HCl (pH ≈ 3.5) for three days, and then freeze-dried to obtain thiol-modified hyaluronic acid with a yield of about 87%, and the thiol modification rate is about 30% as measured by nuclear magnetic resonance.

[0116] Example 24: Preparation of photoresponsive hydrogel A1-B

[0117] Dissolve A1 in water to prepare a 2wt% solution (solution A), dissolve the thiol-modified hyaluronic acid in water to prepare a 2wt% solution (solution B), mix solution A and solution B at a ratio of 1:1, then add 0.1% LAP, mix well, and irradiate the solution with light of wavelength 405 nm for 10-30 s to form a non-flowing hydrogel. Prepare a gel sheet with a diameter of 60 mm and a thickness of 40 mm, place it at the center of the lower plate of the rheometer, apply a pressure of 1 N to the gel block with a 60 mm diameter upper plate, and scan at a fixed frequency (f = 1.592 Hz) in oscillation mode. The elastic modulus of the gel is recorded in Table 1.

[0118] Example 25: Preparation of photoresponsive hydrogel A2-B

[0119] Dissolve A2 in water to prepare a 2wt% solution (solution A), dissolve the thiol-modified hyaluronic acid in water to prepare a 2wt% solution (solution B), mix solution A and solution B at a ratio of 2:1, then add 0.2% LAP, mix well, and irradiate the solution with light of wavelength 405 nm for 10-30 s to form a non-flowing hydrogel. The elastic modulus is measured by rheometer and recorded in Table 1.

[0120] Example 26: Preparation of photoresponsive hydrogel A3-B

[0121] Dissolve A3 in water to prepare a 2wt% solution (solution A), dissolve the thiol-modified hyaluronic acid in water to prepare a 2wt% solution (solution B), mix solution A and solution B at a ratio of 1:1, then add 0.5% LAP, mix well, and irradiate the solution with light of wavelength 405 nm. Within 10-50 s, a non-flowing hydrogel is formed. The elastic modulus is measured by rheometer and recorded in Table 1.

[0122] Example 27: Preparation of photoresponsive hydrogel A4-B

[0123] A4 was dissolved in water to form a 10 wt% solution (solution A), a 10 wt% aqueous solution of dithiothreitol (DTT) was prepared (solution B), solution A and solution B were mixed in a ratio of 4:1, 0.5% I2959 was added, and the solution was mixed uniformly. After the solution was irradiated by light of 365 nm wavelength, a non-flowing hydrogel was formed within 10-50 s. The elastic modulus was measured by a rheometer and recorded in Table 1.

[0124] Example 28: Preparation of photoresponsive hydrogel A5-B

[0125] A5 was dissolved in water to form a 10 wt% solution (solution A), a 20 wt% aqueous solution of polyethylene glycol with two thiol ends (HS-PEG-SH, MW=2000) was prepared (solution B), solution A and solution B were mixed in a ratio of 4:1, 0.5% I2959 was added, and the solution was mixed uniformly. After the solution was irradiated by light of 365 nm wavelength, a non-flowing hydrogel was formed within 10-50 s. The elastic modulus was measured by a rheometer and recorded in Table 1.

[0126] Example 29: Preparation of photoresponsive hydrogel A6-B

[0127] A6 was dissolved in water to form a 2 wt% solution (solution A), a 2 wt% solution of thiol-modified hyaluronic acid was prepared (solution B), solution A and solution B were mixed in a ratio of 1:1, 1% alpha-ketoglutaric acid was added, and the solution was mixed uniformly. After the solution was irradiated by light of 365 nm wavelength for 30 s, a non-flowing hydrogel was formed. The elastic modulus was measured by a rheometer and recorded in Table 1.

[0128] Example 30: Preparation of photoresponsive hydrogel A7-B

[0129] A7 was dissolved in water to form a 2 wt% solution (solution A), a 2 wt% solution of thiol-modified hyaluronic acid was prepared (solution B), solution A and solution B were mixed in a ratio of 1:1, 1% alpha-ketoglutaric acid was added, and the solution was mixed uniformly. After the solution was irradiated by light of 365 nm wavelength for 30 s, a non-flowing hydrogel was formed. The elastic modulus was measured by a rheometer and recorded in Table 1.

[0130] Example 31: Preparation of photoresponsive hydrogel A8-B

[0131] Dissolve A8 in water to make a 2 wt% solution (solution A), dissolve the thiolated modified hyaluronic acid in water to make a 2 wt% solution (solution B), mix solution A and solution B in a 1 : 1 ratio, then add 0.5% I2959, mix well, and irradiate the solution with light of wavelength 365 nm for 20 s to form a non-flowing hydrogel. The elastic modulus was measured by a rheometer and recorded in Table 1.

[0132] Example 32: Preparation of photoresponsive hydrogel A9-B

[0133] Dissolve A9 in water to make a 2 wt% solution (solution A), dissolve the thiolated modified hyaluronic acid in water to make a 2 wt% solution (solution B), mix solution A and solution B in a 1 : 1 ratio, then add 0.1% LAP, mix well, and irradiate the solution with light of wavelength 405 nm for 30 s to form a non-flowing hydrogel. The elastic modulus was measured by a rheometer and recorded in Table 1.

[0134] Example 33: Preparation of photoresponsive hydrogel A10-B

[0135] Dissolve A10 in water to make a 2 wt% solution (solution A), dissolve the thiolated modified hyaluronic acid in water to make a 2 wt% solution (solution B), mix solution A and solution B in a 1 : 1 ratio, then add 0.1% LAP, mix well, and irradiate the solution with light of wavelength 405 nm for 30 s to form a non-flowing hydrogel. The elastic modulus was measured by a rheometer and recorded in Table 1.

[0136] Example 34: Preparation of photoresponsive hydrogel A11-B

[0137] Dissolve A11 in water to make a 2 wt% solution (solution A), dissolve the thiolated modified hyaluronic acid in water to make a 2 wt% solution (solution B), mix solution A and solution B in a 1 : 1 ratio, then add 0.1% LAP, mix well, and irradiate the solution with light of wavelength 405 nm for 30 s to form a non-flowing hydrogel. The elastic modulus was measured by a rheometer and recorded in Table 1.

[0138] Example 35: Preparation of photoresponsive hydrogel A12-B

[0139] Dissolve A12 in water to make a 2 wt% solution (solution A), dissolve the thiolated modified hyaluronic acid in water to make a 2 wt% solution (solution B), mix solution A and solution B in a 1 : 1 ratio, then add 0.1% LAP, mix well, and irradiate the solution with light of wavelength 405 nm for 30 s to form a non-flowing hydrogel. The elastic modulus was measured by a rheometer and recorded in Table 1.

[0140] Example 36: Preparation of photoresponsive hydrogel A13-B

[0141] A13 was dissolved in water to prepare a 2 wt% solution (solution A), and the thiolated modified hyaluronic acid was dissolved in water to prepare a 2 wt% solution (solution B). Solution A and solution B were mixed at a ratio of 1 : 1, and 0.1% LAP was added. The solution was irradiated with light at a wavelength of 405 nm for 30 s to form a non-flowing hydrogel. The elastic modulus was measured by a rheometer and recorded in Table 1.

[0142] Example 37: Preparation of photoresponsive hydrogel A14-B

[0143] A14 was dissolved in water to prepare a 10 wt% solution (solution A), and the thiolated modified hyaluronic acid was dissolved in water to prepare a 2 wt% solution (solution B). Solution A and solution B were mixed at a ratio of 3: 1, and 0.1% LAP was added. The solution was irradiated with light at a wavelength of 405 nm for 30 s to form a non-flowing hydrogel. The elastic modulus was measured by a rheometer and recorded in Table 1.

[0144] Example 38: Preparation of photoresponsive hydrogel A15-B

[0145] A15 was dissolved in water to prepare a 10 wt% solution (solution A), and a 10 wt% dithiothreitol (DTT) aqueous solution (solution B) was prepared. Solution A and solution B were mixed at a ratio of 4: 1, and 0.1% LAP was added. The solution was irradiated with light at a wavelength of 405 nm for 30 s to form a non-flowing hydrogel. The elastic modulus was measured by a rheometer and recorded in Table 1.

[0146] Example 39: Preparation of photoresponsive hydrogels A16-B-1 and A16-B-2

[0147] A16 and a16 were dissolved in water to prepare a 2 wt% solution (solution A and solution a, respectively), and the thiolated modified hyaluronic acid was dissolved in water to prepare a 2 wt% solution (solution B). Solution A and solution B were mixed at a ratio of 1 : 1, and 0.2% LAP was added. The solution was irradiated with light at a wavelength of 405 nm for 20 s to form a non-flowing hydrogel A16-B-1. Solution a and solution B were mixed at a ratio of 1 : 1, and 0.2% LAP was added. The solution was irradiated with light at a wavelength of 405 nm for 20 s to form a non-flowing hydrogel A16-B-2. The elastic modulus was measured by a rheometer and recorded in Table 1.

[0148] Examples 40-45: Preparation of photoresponsive hydrogels A17-B to A22-B

[0149] The polymer A16 was replaced by A17, A18, A19, A20, A21, A22 in turn in the method as described in Example 39, and the remaining steps were unchanged to obtain different combination solutions. After irradiation for 10-30 s, the non-flowing hydrogels A17-B~A22-B were formed. The elastic modulus was recorded in Table 1 by a rheometer.

[0150] Table 1 Composition and gel performance data of light-responsive hydrogels A1-B~A22-B

[0151] Photoresponsive hydrogel Solution A, concentration Solution B, concentration Photoinitiator, concentration Photocuring time Gel elastic modulus A1-B A1, 1% HASH, 1% LAP, 0.1% 15s 45 kPa A2-B A2, 1.33% HASH, 0.67% LAP, 0.2% 20s 33 kPa A3-B A3, 1% HASH, 1% LAP, 0.5% 15s 47 kPa A4-B A4, 8% DTT, 2% I2959, 0.5% 30s 53 kPa A5-B A5, 8% SH-PEG 2000 -SH, 4%]]> I2959, 0.5% 30s 46 kPa A6-B A6, 1% HASH, 1% Alpha-ketoglutaric acid, 1% 30s 40 kPa A7-B A7, 1% HASH, 1% Alpha-ketoglutaric acid, 1% 30s 45 kPa A8-B A8, 1% HASH, 1% I2959, 0.5% 20s 36 kPa A9-B A9, 1% HASH, 1% LAP, 0.1% 30s 39 kPa A10-B A10, 1% HASH, 1% LAP, 0.1% 30s 47 kPa A11-B A11, 1% HASH, 1% LAP, 0.1% 30s 38 kPa A12-B A12, 1% HASH, 1% LAP, 0.1% 30s 36 kPa A13-B A13, 1% HASH, 1% LAP, 0.1% 30s 38 kPa A14-B A14, 7.5% HASH, 0.5% LAP, 0.1% 30s 38 kPa A15-B A15, 8% DTT, 2% LAP, 0.1% 30s 37 kPa A16-B-1 A16, 1% HASH, 1% LAP, 0.2% 20s 39 kPa A16-B-2 a16, 1% HASH, 1% LAP, 0.2% 20s 41 kPa A17-B A17, 1% HASH, 1% LAP, 0.2% 20s 38 kPa A18-B A18, 1% HASH, 1% LAP, 0.2% 20s 37 kPa A19-B A19, 1% HASH, 1% LAP, 0.2% 20s 40 kPa A20-B A20, 1% HASH, 1% LAP, 0.2% 20s 36kPa A21-B A21, 1% HASH, 1% LAP, 0.2% 20s 34 kPa A22-B A22, 1% HASH, 1% LAP, 0.2% 20s 39 kPa

[0152] Example 46: Evaluation of sunscreen effect by measuring the ultraviolet absorbance or transmittance of light-responsive hydrogels

[0153] The hydrogel precursor solutions prepared in Examples 24-32 were spread on polymethyl methacrylate plates with a thickness of about 1 mm, and the sunscreen factors (sunscreen factor 1) of each group of samples were tested by a UV transmittance analyzer. Then each group of samples was irradiated by blue light with a wavelength of 405 nm for 60 s, and then immersed in water for 20 min. After taking out, the sunscreen factors (sunscreen factor 2) were tested again to evaluate the sunscreen performance of each group of sunscreen products. The 0.1% p-nitro-cinnamic acid DMF solution and the 0.1% benzoic acid DMF solution were used as Comparative Example 1 and Comparative Example 2, respectively. The test results are shown in Table 2, which is the sunscreen factor detection data table of the light-responsive hydrogels A1-B~A9-B prepared in Examples 24-32. The data show that based on the natural polymer derivative having the structure shown in Formula I, the carbon-carbon double bond and the sunscreen functional group are introduced into the backbone of the natural polymer derivative having the structure shown in Formula I. The light-responsive hydrogel comprises a natural polymer derivative having the structure shown in Formula I (component A), a crosslinking agent containing multiple thiol groups (component B), and a photoinitiator. The two components A and B can undergo crosslinking reaction to form a hydrogel under the action of the photoinitiator after irradiation. At this time, the sunscreen functional group on the natural polymer derivative having the structure shown in Formula I can enhance the sunscreen function at the application site. At the same time, the cinnamic acid or cinnamic acid derivative; the cinnamic acid derivative will undergo photodimerization to form a crosslinked network under specific wavelength irradiation, which can improve the mechanical properties of the light-responsive hydrogel. That is, the crosslinked network can increase the residence time of the sunscreen product and prolong the period of sunscreen efficacy.

[0154] Table 2

[0155] Photoresponsive hydrogel Sun Protection Factor 1 Sun Protection Factor 2 A1-B 1550 1549 A2-B 1674 1658 A3-B 1521 1530 A4-B 1537 1533 A5-B 1602 1592 A6-B 1564 1561 A7-B 1543 1540 A8-B 1582 1574 A9-B 1589 1578 Comparative Example 1 1533 1046 Comparative Example 2 1572 1083

[0156] Example 47: Tissue adhesion strength test and burst pressure test

[0157] For Examples 33-38, after cleaning excess grease from fresh pig casings, cut them into 4cm × 2cm pieces and fix them onto a 6.5cm × 2.5cm glass slide using 502 glue. Evenly apply 0.2 mL of hydrogel precursor solution to the surface of the casing. Then, place another glass slide on top of this slide and irradiate the casing area with a 405nm LED light source for 5 minutes to allow the hydrogel precursor solution to gel in situ between the two casing pieces. After complete gelation, fix one end of the glass slide vertically, connect the other end to a digital tensile tester, and slowly pull it horizontally. Figure 1 As shown, the maximum tensile force when the two sausage casings separate is recorded and divided by the contact area of ​​the two sausage casings to obtain the tissue adhesion strength of the hydrogel.

[0158] After cleaning fresh pig intestines, cut them into 4×4 cm pieces. Cover the burst pressure testing device with the intestines and cut a 2 mm diameter circular hole in the intestine. Cover the cut with 0.5 mL of hydrogel precursor solution and irradiate with 405 nm blue light for 60 seconds to form hydrogel in situ at the puncture site. The hydrogel thickness is approximately 4.4 mm. Measure the burst pressure after gelation. Pour water into the device; the peak pressure before pressure loss is the burst pressure. Figure 2 As shown. A commercially available sample, Fibrin Glue (manufacturer Baxter, model TISSEEL), was used as a comparison, and all measurements were repeated three times. The tissue adhesion and burst pressure of the hydrogels are shown in Table 3, which is a data table of tissue adhesion and burst pressure tests for the photoresponsive hydrogels A10-B~A15-B prepared in Examples 33-38. Data shows that, based on the natural polymer derivative with the structure shown in Formula I of this invention, carbon-carbon double bonds and small molecule functional groups containing polyphenols are introduced into its backbone. The photoresponsive hydrogel includes a natural polymer derivative with the structure shown in Formula I (component A), a crosslinking agent containing multiple thiol groups (component B), and a photoinitiator. Components A and B can undergo a crosslinking reaction to form a hydrogel after being exposed to light under the action of the photoinitiator. The relative proportions of its tissue adhesion strength and burst pressure performance are improved, making it more advantageous for use in antibacterial, antioxidant, and wet tissue adhesion products. In other words, the small molecule functional groups containing polyphenols on the natural polymer derivative with the structure shown in Formula I can enhance the antibacterial, antioxidant, and wet tissue adhesion effects at the application site.

[0159] Table 3

[0160] Photoresponsive hydrogel Tissue adhesion strength (kPa) Burst pressure (kPa) A10-B 23.7 28.1 A11-B 24.3 27.5 A12-B 25.2 29.8 A13-B 32.1 38.8 A14-B 33.8 39.3 A15-B 21.7 30.3 Fibrin Glue 17.5 22.4

[0161] Example 48: Hydrogel-Promoted Skin Repair Test

[0162] A 1.5 cm x 1.5 cm skin defect was created on the back of a mature female SD rat using surgical scissors to establish a rat skin defect model. The wound surface was covered with 0.5 mL of the hydrogel precursor solution of A16-B-1 and A16-B-2 (without EFG short peptide modification) in Example 39, respectively, and irradiated with 405 nm blue light for 60 s to form a hydrogel in situ at the defect site. After 2 weeks of treatment and observation, the wound closure rate of the mice in the A16-B-1 group was 91%, the wound closure rate of the mice in the blank control group without treatment was 46%, and the wound closure rate of the mice in the positive control group covered with A16-B-2 hydrogel was 65% (as shown in Figure 3 , the results show that the EGF short peptide modified hydrogel A16-B-1 has a significant effect on promoting tissue repair. It is shown that the small molecule functional group containing a polyphenol structure on the natural polymer derivative of Formula I can enhance its function at the application site.

[0163] Example 49 Hydrogel promotes skin repair test

[0164] A 1.5 cm x 1.5 cm skin defect was created on the back of a mature female SD rat using surgical scissors to establish a rat skin defect model. The wound surface was covered with 0.5 mL of the hydrogel precursor solution of A20-B in Example 42 and A16-B-2 (without EFG short peptide modification) in Example 39, respectively, and irradiated with 405 nm blue light for 60 s to form a hydrogel in situ at the defect site. After 2 weeks of treatment and observation, the wound closure rate of the mice in the A20-B group was 84%, the wound closure rate of the mice in the blank control group without treatment was 46%, and the wound closure rate of the mice in the positive control group covered with A16-B-2 was 65% (as shown in Figure 3 , the results show that the RGD peptide modified hydrogel has a significant effect on promoting tissue repair. It is further shown that the small molecule functional group containing a polyphenol structure on the natural polymer derivative of Formula I can enhance its function at the application site.

[0165] The technical features of the above-described embodiments can be combined in any manner. To make the description concise, not all possible combinations of the technical features in the above-described embodiments are described, but as long as the combinations of the technical features do not contradict each other, they should be considered within the scope of the present disclosure.

[0166] The above-described embodiments only express several embodiments of the present application, and the description is more specific and detailed, but it should not be construed as limiting the scope of the patent. It should be noted that for those skilled in the art, without departing from the concept of the present application, a number of modifications and improvements can be made, which are within the scope of the present application. Therefore, the scope of the patent of the present application should be subject to the appended claims.

Claims

1. A multifunctional light-responsive hydrogel, characterized in that, Prepared from component A, component B and a photoinitiator in a solvent, wherein component A is a natural polymer derivative, component B is a crosslinking agent containing multiple mercapto groups; the concentration of component A is 1-5wt%, the concentration of component B is 1-5wt%, the concentration of the photoinitiator is 0.001-1wt%, and the rest is the solvent; The crosslinking agent containing multiple mercapto groups is selected from any one of dithiothreitol, mercapto-polyethylene glycol or mercapto-modified hyaluronic acid, gelatin, collagen, alginic acid, chitosan; The natural polymer derivative has a structure as shown in formula I: ， In formula I, P is a natural polymer backbone, 1、 P2 is independently selected from natural polymer repeat units, R1 is a carbon-carbon double bond-containing group; R2 is a UV-absorbing small molecule, including at least one of cinnamic acid, cinnamic acid derivatives, X, Y are each independently selected from an ether bond, an ester bond, an amide bond, a coupling molecule, the coupling molecule at least includes two grafting sites of amino and / or hydrazine, one end is connected to the carboxyl site on the natural polymer through the amidation reaction, and the other end is connected to R1 or R2; m, n are each independently selected from an integer of 1-10000; The natural polymer includes at least one of sodium hyaluronate, sodium alginate, chitosan, gelatin, collagen, chondroitin sulfate or fibrin; The preparation method of the natural polymer derivative selects any one of the following preparation methods a-d: a. R1 and R2 are sequentially modified on the hydroxyl, carboxyl or amino sites of the natural polymer; b. A coupling molecule containing amino and / or hydrazide is introduced on the carboxyl site of the natural polymer, and then R1 and R2 are sequentially modified on the amino and / or hydrazide; c. R1 is modified on the carboxyl or hydroxyl site of the natural polymer, a coupling agent containing amino is introduced on the carboxyl site of the natural polymer derivative modified by R1, and then R2 is modified on the amino; d. R2 is modified on the carboxyl or hydroxyl site of the natural polymer, a coupling agent containing amino and / or hydrazide is introduced on the carboxyl site of the natural polymer derivative modified by R2, and then R1 is modified on the amino or hydrazide.

2. The multifunctional light-responsive hydrogel according to claim 1, wherein The group containing carbon-carbon double bond is selected from vinyl, propenyl, allyloxy, cyclohexenyl, norbornenyl, bicyclo[2.2.2]oct-5-enyl.

3. The multifunctional light-responsive hydrogel according to claim 1, wherein The photoinitiator includes at least one of lithium phenyl(2,4,6-trimethylbenzoyl)phosphinate, 2-hydroxy-2-methyl-1-[4-(2-hydroxyethoxy)phenyl]-1-propanone, eosin Y, alpha-ketoglutaric acid, 2,4,6-trimethylbenzoyl diphenyl phosphine oxide or 1-hydroxycyclohexyl benzophenone.

4. The method for preparing a multifunctional photo-responsive hydrogel according to claim 1, wherein, The method includes the following steps: Components A, component B and photoinitiator are dissolved in a solvent to obtain a hydrogel precursor solution: The hydrogel precursor solution is irradiated under a light source to undergo photocrosslinking to form a hydrogel.

5. The use of the multifunctional photo-responsive hydrogel according to claim 1, wherein, Selected from the following applications: The application of the light-responsive hydrogel in ultraviolet protection products.

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