Preparation and use of hydrogel

Abstract

Provided are the preparation and use of a hydrogel. The hydrogel is composed of two polymers A and B, wherein the main chains of the two polymers A and B are composed of amide bonds formed by means of the polycondensation of lysine and glutamic acid, and the value of n is 8-100; and the two polymers A and B are each a polymer having any branching generation. The structural formulae of the two polymers A and B are respectively represented by formulae (a) and (b): each Ra on the main chains of the two polymers A and B is selected from a structure as represented by formula (c) or (d), wherein in the formulae, Ra1 is an exogenous functional molecule connected to the tail ends of alkynyl groups on the main chains of the two polymers A and B; Rb is a branched structure at the tail ends of amino groups in side chains on the skeletons of the two polymers A and B; Rc is a monosaccharide or oligosaccharide connected to the tail end of Rb of the polymer A; and Rd is any phenylboronic acid structure compound connected to the tail end of Rb of the polymer B. The hydrogel has a relatively good biological safety and can be used in the preparation of medical preparations, etc.

Description

Preparation and use of hydrogel Technical Field

[0001] The invention belongs to the field of biomaterials and biomedicine, and particularly relates to the preparation and application of a hydrogel. Background Art

[0002] Hydrogels are a type of three-dimensional network composed of hydrophilic polymer chains that are cross-linked to form a matrix with a high water content (Annabi N., Tamayol A., Uquillas JA, et al. 25th Anniversary Article: Rational Design and Applications of Hydrogels in Regenerative Medicine [J]. Advanced Materials, 2014, 26(1): 85-124.). Because the polymer backbone in the hydrogel has a hydrophilic part, the hydrogel material can retain a large amount of water inside, so it has similar physical and chemical properties to liquid water, while exhibiting macroscopic solid-like rheological behavior (Guan Q.-F., Yang H.-B., Han Z.-M., et al. Sustainable Cellulose-Nanofiber-Based Hydrogels[J]. ACS Nano, 2021, 15(5): 7889-7898 & Davidson MD, Ban E., Schoonen ACM, et al. Mechanochemical Adhesion and Plasticity in Multifiber Hydrogel Networks[J]. Advanced Materials, 2020, 32(8): 1905719.). Hydrogels have become advanced materials for novel applications, ranging from drug delivery to wound healing, contact lenses for enhanced vision, food, tissue engineering, and many other technological applications. Many parts of the human body contain hydrogels in the form of extracellular matrix, collagen, mucus, gelatin, cartilage, menisci, epidermis, vitreous humor, and tendons. The hydrophilicity and 3D structure of hydrogels give them the ability to retain large amounts of water or biological fluids. Hydrogels are composed of dynamic cross-linked structures, which enable them to maintain the integrity of the hydrogel network.This property of hydrogels enables them to mimic many properties of the extracellular matrix in tissues (Brown TE, Anseth KSSpatiotemporal hydrogel biomaterials for regenerative medicine[J].Chemical Society Reviews,2017,46(21):6532-6552&Thiele J.,Ma Y.,Bruekers SMC,et al.25th Anniversary Article:Designer Hydrogels for Cell Cultures:A Materials Selection Guide[J].Advanced Materials,2014,26(1):125-148.), and they are widely used in tissue engineering and biomedical applications.

[0003] Macroscopic hydrogel materials formed by sugar-based macromolecules can be used in tissue engineering applications. The natural extracellular matrix (ECM) is composed of a complex network of biomacromolecules surrounding cells. On the one hand, ECM provides different mechanical properties in different tissues, which is very important for stabilizing tissue structure and mechanics; on the other hand, ECM has a complex macromolecular composition and can provide many biologically active signals to the cells surrounded by it. The characteristics of ECM in both mechanical properties and biochemical signals play an important role in regulating various cell behaviors (Frantz C., Stewart KM, Weaver VM The extracellular matrix at a glance [J]. Journal of Cell Science, 2010, 123 (24): 4195-4200.). Sugar-based macromolecular materials have special advantages as tissue engineering scaffolds. They can not only provide adjustable mechanical properties by virtue of their rich chemical structure and supramolecular interactions, but also provide biochemical signals that mediate cell-scaffold interactions as bioactive molecules. Therefore, sugar-based macromolecular materials can act as ideal ECM mimics to construct biomaterial scaffolds to regulate related cell behaviors and promote tissue regeneration (O'Brien FJ Biomaterials & scaffolds for tissue engineering [J]. Materials Today, 2011, 14 (3): 88-95 & Place ES, Evans ND, Stevens MM Complexity in biomaterials for tissue engineering [J]. Nature Materials, 2009, 8 (6): 457-470.).

[0004] Traditional hydrogels use synthetic polymers as raw materials, such as polyethylene glycol, polyvinyl alcohol, poly (2-hydroxyethyl methacrylate) and polyacrylamide, and are cross-linked through chemical reactions. These hydrogels generally exhibit high mechanical properties and have certain degradation capabilities. However, traditional hydrogels generally lack biological activity and have limited biocompatibility. The covalent cross-linking method makes them lack dynamic adaptability to cells, and they generally have low adhesion strength to wet surfaces and biological tissues (Zhao Y., Song S., Ren X., et al. Supramolecular Adhesive Hydrogels for Tissue Engineering Applications [J]. Chemical Reviews, 2022, 122 (6): 5604-5640.). Various hydrogels based on alginate, collagen, agarose, hyaluronic acid and chitosan can be prepared by physical cross-linking methods using natural biomacromolecules. These hydrogels have better biocompatibility and certain biological activity. However, the lack of sufficient mechanical properties of these materials greatly limits their application range (Li Y., Rodrigues J., Tomás H. Injectable and biodegradable hydrogels: gelation, biodegradation and biomedical applications [J]. Chemical Society Reviews, 2012, 41(6): 2193-2221.).

[0005] Mooney et al. have developed a dual-network polymer hydrogel that simultaneously forms an ionic interaction crosslinking and a covalently crosslinked network. The strength of the gel has been significantly improved, and it can withstand deformation exceeding 20 times its initial length, and the fracture energy is greatly improved. However, the presence of polyacrylamide and covalent crosslinking in the gel network still reduces its degradation performance and dynamic stress relaxation performance (Sun J.-Y., Zhao X., Illeperuma WRK, et al. Highly stretchable and tough hydrogels [J]. Nature, 2012, 489 (7414): 133-136.). The scope of the modulus of existing supramolecular hydrogels is generally narrow, and the strength difference of different tissues is very large. The modulus range of supramolecular hydrogels cannot meet the strength requirements of tissue repair.

[0006] With the continuous development of science and technology and medical level, as well as people's increasing requirements for the quality and efficiency of medical treatment, there are more and more types of medical adhesives, which can be mainly divided into two categories, namely soft tissue adhesives and hard tissue adhesives. Soft tissue adhesives can be used for bonding of skin, organs, nerves, blood vessels, mucous membranes and other parts. They mostly use medical α-cyanoacrylate glue and fibrin bio-glue. Since fibrin bio-glue is produced in the blood, there is a possibility of mutual transmission of infectious diseases such as hepatitis and AIDS during use. Although autologous blood can be used, it is not suitable for emergency treatment, and its strength and speed are not ideal. Among them, ophthalmic surgery is the fundamental clinical means to solve a variety of eye diseases. Suture suturing is currently the standard method for repairing eye incisions, but it is prone to cause postoperative complications such as irritation and inflammation. In order to overcome the shortcomings of traditional sutures, medical gel adhesives have gradually been developed in recent years. Commonly used adhesives include (Baxter AG), However, they all have limitations such as poor biocompatibility, weak adhesion strength or the need for low-temperature storage, and cannot meet the requirements of actual clinical applications.

[0007] Summary of the Invention

[0008] In a first aspect, the present invention provides a hydrogel, comprising two polymers, A and B, wherein the main chains of the two polymers A and B are composed of amide bonds formed by condensation polymerization of lysine and glutamic acid, and the value of n is 8-100; the two polymers A and B are polymers having any number of branching generations; the structural formulas of the two polymers A and B are shown as a and b, respectively:

[0009] Ra on the main chains of the two polymers A and B is selected from (c) or (d), and the structural formulas are shown in (c) and (d):

[0010] or

[0011] Where R a1 Connecting exogenous functional molecules to the alkyne ends of the two polymer main chains A and B; R b It is the branched structure of the amino terminal of the side chain on the two polymer backbones A and B; R c R of polymer A b Terminally linked monosaccharide or oligosaccharide; R d R of polymer B b Any phenylboronic acid structure compound connected at the end.

[0012] Furthermore, the functional molecules are selected from one or more of peptides, fluorescent groups, antibacterial molecules, growth factors, functional oligosaccharides or polysaccharides.

[0013] Furthermore, the peptide segment is selected from cell adhesive peptides, antimicrobial peptides, hormone peptides, immunomodulatory peptides, and antioxidant peptides, and the functional oligosaccharide or polysaccharide can be one or more of glycosaminoglycans, fucoidan, alginate, hyaluronic acid, and chitosan.

[0014] Furthermore, the R b The branched structure is a branching reaction that can be carried out on the amino ends of the side chains of polymers A and B.

[0015] Furthermore, the branching generations are preferably 0-5 generations.

[0016] Furthermore, the branching of the amino terminal is performed by using a compound containing an amino group to increase the number of amino terminals.

[0017] Furthermore, the branching method is selected from the branching method using a compound containing at least two amino groups and one carboxyl group or a branching method using a substance that forms polyamide-amine.

[0018] Furthermore, the compound containing an amino group is selected from one or more of lysine, diaminopropionic acid, diaminobutyric acid, diaminobutyric acid, diaminohexanoic acid, and diaminoheptanoic acid.

[0019] Furthermore, the amino termini of the branched structures of the polymer A can be connected to different types of monosaccharides or oligosaccharides.

[0020] Furthermore, the monosaccharide can be selected from one or more of mannose, glucose, galactose, fructose, xylose, fucose, gulose, idose, lyxose, mannuronic acid, glucuronic acid, galacturonic acid, guluronic acid, and derivatives thereof.

[0021] Furthermore, the oligosaccharide may be one or more of any oligosaccharide.

[0022] Furthermore, the amino terminal of the branched structure of the polymer B is connected to any phenylboronic acid structure, including one or more of phenylboronic acid half ester and carbonylphenylboronic acid half ester.

[0023] Furthermore, the polymers A and B in the hydrogel are mixed in a solution with a solid content of 10%-20%.

[0024] Furthermore, the ratio of polymers A and B in the hydrogel is 1-2:2-1.

[0025] Furthermore, the strength of the hydrogel is 10Pa-100000Pa.

[0026] Furthermore, the hydrogel has excellent tissue adhesion, shear resistance, visible light transmittance and / or biodegradability.

[0027] In a second aspect, the present invention provides use of the hydrogel described in the first aspect in preparing skin wound material.

[0028] Furthermore, the skin trauma refers to trauma to the skin or mucous membrane caused by burns, mechanical damage or inflammation, and the mucous membrane includes oral mucosa or nasal mucosa, etc.

[0029] Furthermore, the substance for repairing wounds may be a drug or a medical device.

[0030] In a third aspect, the present invention provides use of the hydrogel described in the first aspect in preparing a suture required for conjunctival transplantation or corneal transplantation.

[0031] Furthermore, the use may be to improve the adhesion of conjunctival transplantation or corneal transplantation in situ and the closure of wounds.

[0032] Furthermore, the invention can be used as a suture agent to replace sutures.

[0033] In a fourth aspect, the present invention provides a medical preparation comprising the hydrogel according to the first aspect of the present invention.

[0034] Furthermore, the preparation also contains anti-inflammatory, hemostatic and other preparations and pharmaceutical excipients.

[0035] In a fifth aspect, the present invention provides a use of the hydrogel described in the first aspect in preparing a bone repair material.

[0036] Furthermore, the bone repair includes but is not limited to trauma, spine, joint, orthopedics, etc.

[0037] In a sixth aspect, the present invention provides a method for preparing the hydrogel according to the first aspect:

[0038] S1. Preparation of intermediate 1:

[0039] The raw materials Boc-Lys(Boc)-OH and H-Glc(OMe)-OMe·HCl underwent condensation reaction with the reagents EDC, HOBT, and DIPEA to obtain the intermediate product 1.

[0040] S2. Preparation of intermediate product 2:

[0041] Intermediate 1 undergoes deprotection reaction in the presence of hydrogen chloride ethyl acetate solution to obtain intermediate 2.

[0042] S3. Preparation of intermediate product 3:

[0043] Intermediate 2 and the raw material Boc-Lys(Boc)-OH underwent condensation reaction in the presence of reagents EDC, HOBT, and DIPEA to obtain intermediate 3.

[0044] S4. Preparation of intermediate products 4 and 5:

[0045] Intermediate product 1 or 3 undergoes deprotection reaction in the presence of lithium hydroxide to obtain intermediate product 4 or 5, respectively.

[0046] S5. Preparation of intermediates 6, 7, and 8:

[0047] The raw material Boc-Glc-OH or intermediate product 4 or 5 and the raw material pentafluorophenol undergo condensation reaction under the action of the reagent EDC to obtain intermediate products 6, 7, and 8, respectively.

[0048] S6. Preparation of intermediate product 9:

[0049] The raw material Boc-Lys(Boc)-OH and propargylamine reacted with the reagents EDC, HOBT, and DIPEA to obtain the intermediate product 9.

[0050] S7. Preparation of intermediate product 10 (Mb):

[0051] Intermediate product 9 undergoes deprotection reaction in the presence of hydrogen chloride ethyl acetate solution to obtain intermediate product 10 (Mb).

[0052] S8. Preparation of intermediate product 11 (Mc):

[0053] The raw materials Boc-aminoacetic acid and pentafluorophenol underwent a condensation reaction with the reagent EDC to obtain the intermediate 11 (Mc).

[0054] S9. Preparation of intermediates 12, 13, and 14:

[0055] Intermediate products 6, 7, or 8 and intermediate product 10 (Mb) undergo polymerization reaction under the action of reagent DIPEA to obtain intermediate products 12, 13, and 14.

[0056] S10. Preparation of intermediates 15, 16, and 17:

[0057] Intermediate 12, 13, or 14 undergoes deprotection reaction under the action of trifluoroacetic acid to obtain intermediates 15, 16, and 17, respectively.

[0058] Preparation of S11.B component (products 18, 19, 20):

[0059] Intermediate product 15, 16, or 17 reacts with raw material Bob in the presence of sodium borohydride to obtain component B (products 18, 19, and 20), respectively.

[0060] S12. Preparation of intermediates 21, 22, and 23:

[0061] Intermediate 15, 16, or 17 reacts with intermediate 11 (Mc) in the presence of DIPEA, and further undergoes deprotection in the presence of trifluoroacetic acid to afford intermediates 21, 22, and 23, respectively.

[0062] Preparation of S13.A component (products 24, 25, 26):

[0063] The intermediate product 21, 22, or 23 and the raw material monosaccharide or oligosaccharide undergo condensation reaction under the action of the reagent aniline to obtain the product A component (product 24, 25, 26) respectively;

[0064] S14. Synthesis of Hydrogel

[0065] The solid content of component A or component B is prepared at 12 wt% to 20 wt%. BRIEF DESCRIPTION OF THE DRAWINGS

[0066] Figure 1 Molecular structures of raw material A and B components

[0067] Figure 2 Changes in branching structure in group A / B

[0068] Figure 3 Synthesis routes of intermediates M0-a, M1-a, M2-a, Mb, and Mc

[0069] Figure 4 1 H NMR result of 1

[0070] Figure 5 1 H NMR result of 2

[0071] Figure 61H NMR result of 3

[0072] Figure 7 1 H NMR result of 4

[0073] Figure 8 1 H NMR result of 5

[0074] Figure 9 1 H NMR result of 6

[0075] Figure 10 1 H NMR result of 7

[0076] Figure 11 1 H NMR result of 8

[0077] Figure 12 1 H NMR result of 9

[0078] Figure 13 1 H NMR result of 10

[0079] Figure 14 1 H NMR result of 11

[0080] Figure 15 Synthesis routes of P0, P1, P2, component A (P0-Man(Glc), P1-Man(Glc), P2-Man(Glc)), and component B (P0-Bob, P1-Bob, P2-Bob)

[0081] Figure 16 1 H NMR result of 12

[0082] Figure 17 1 H NMR result of 15

[0083] Figure 18 1 H NMR result of compound 16

[0084] Figure 19 1 H NMR result of compound 17

[0085] Figure 20 1 H NMR result of 18

[0086] Figure 21 1 H NMR result of 19

[0087] Figure 22 1 H NMR result of 20

[0088] Figure 23 1 H NMR result of 21

[0089] Figure 24 1 H NMR result of 22

[0090] Figure 25 1 H NMR result of 23

[0091] Figure 26 1H NMR result of 24(P0-Man)

[0092] Figure 27 1 H NMR result of 25(P1-Man)

[0093] Figure 28 1 H NMR result of 25(P1-Glc)

[0094] Figure 29 1 H NMR result of 26(P2-Man)

[0095] Figure 30 Synthesis routes of polymers P0-Bob-1, P1-Bob-1, and P2-Bob-1 with replacement of phenylboronic acid ester structures

[0096] Figure 31 1 H NMR result of P1-Bob-1

[0097] Figure 32 1 H NMR result of 30(P2-Man-RGD)

[0098] Figure 33 Synthesis routes of polymers G0, G1, and G2 with varying branching patterns

[0099] Figure 34 1 H NMR result of G0.5

[0100] Figure 35 1 H NMR result of G1.0

[0101] Figure 36 1 H NMR result of G1.5

[0102] Figure 37 1 H NMR result of G2.0

[0103] Figure 38 Tissue Adhesion Strength ((a) P0-Man+P0-Bob, P1-Man+P1-Bob, P2-Man+P2-Bob; (b) P2-Man+P0-Bob, P2-Man+P0-Bob, P2-Man+P2-Bob; (c) P1-Bob+P1-Glc, P1-Bob+P1-Man)

[0104] Figure 39 Shear resistance test

[0105] Figure 40 Mechanical strength of hyperbranched polypeptide hydrogels constructed under the same branching system conditions with a solid content of 12 wt% (a) P0-Man+P0-Bob (b) P1-Man+P1-Bob (c) P2-Man+P2-Bob

[0106] Figure 41 Mechanical strength of hyperbranched polypeptide hydrogels constructed under different branching system conditions with a solid content of 12 wt% (a) P2-Man + P2-Bob (b) P2-Man + P1-Bob (c) P0-Man + P0-Bob (d) P1-Man + P0-Bob (e) P2-Man + P0-Bob

[0107] Figure 42 Mechanical strength of gels formed by P1-Glc+P1-Bob and P1-Man+P1-Bob at neutral conditions

[0108] Figure 43 Hydrogel adhesion stability test on pig skin in vitro under external force

[0109] Figure 44 Underwater test of gel adhesion stability test

[0110] Figure 45 Light transmittance test

[0111] Figure 46 In vitro degradation experiment

[0112] Figure 47 Gel autologous conjunctival transplantation test

[0113] Figure 48 Biomechanical test of hydrogel sealing corneal incision in pig eyes in vitro

[0114] Figure 49 Biosafety Test

[0115] Figure 50 Cell safety test

[0116] Figure 51 Rabbit autologous conjunctival transplantation experiment

[0117] Figure 52 Corneal damage repair effect in an animal model of corneal damage repair

[0118] Figure 53 Bone defect repair effect in animal model of bone defect repair DETAILED DESCRIPTION

[0119] Example 1 Synthesis of hydrogel pre-materials

[0120] In this embodiment, the raw materials required for hydrogel synthesis include Boc-L-glutamic acid (Boc-Glc-OH), (S)-2,6-di-tert-butyloxycarbonylaminohexanoic acid (Boc-Lys(Boc)-OH), L-glutamic acid dimethyl ester hydrochloride (H-Glc(OMe)-OMe·HCl), pentafluorophenol, mannose (or different types of monosaccharides or oligosaccharides), phenylboronic acid half ester (Bob), Boc-aminooxyacetic acid, and propargylamine; all of which are commercially available.

[0121] The required reagents include 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC), 1-hydroxybenzotriazole (HOBT), N,N-diisopropylethylamine (DIPEA), lithium hydroxide, hydrogen chloride ethyl acetate solution, sodium borohydride, trifluoroacetic acid, anhydrous ether, and aniline, all of which are commercially available.

[0122] 1.1 Molecular structure

[0123] 1.1.1 Basic molecular structure of raw materials

[0124] The structure diagram of raw material A component and raw material B component is shown in Figure 1.

[0125] 1.2 Molecular expansion structure

[0126] 1.2.1 The changes in branching structure of group A / B are shown in Figure 2.

[0127] 1.2.2 Changes in sugar content in component A

[0128] R=mannose (Man) Or R=glucose (Glc)

[0129] 1.3 Preparation and characterization of components A and B

[0130] 1.3.1 Preparation of intermediates M0-a, M1-a, M2-a, Mb, and Mc (see Figure 3);

[0131] 1. Synthesis of Compound 1

[0132] (1) Boc-Lys(Boc)-OH, EDC, HOBT and H-Glc(OMe)-OMe·HCl were dissolved in dry dichloromethane.

[0133] (2) Slowly add DIPEA or triethylamine dropwise in an ice-water bath. The molar ratio of the raw materials is Boc-Lys(Boc)-OH:EDC:HOBT:H-Glc(OMe)-OMe·HCl:DIPEA or triethylamine = 1:1:1:1:1.

[0134] (3) Remove the ice-water bath and continue stirring for 6 to 36 hours. After the reaction is completed, wash with dilute hydrochloric acid, dilute sodium hydroxide, and saturated brine. After separating the organic phase, the crude product is separated and purified by silica gel column chromatography to obtain a white solid (see Figure 4).

[0135] 2. Synthesis of Compound 2

[0136] (1) In an ice-water bath, an excess of saturated hydrogen chloride ethyl acetate solution was slowly added dropwise to compound 1.

[0137] (2) Remove the ice-water bath and stir the reaction for 30 minutes. A large amount of white precipitate will be produced during the reaction.

[0138] (3) After the reaction is completed, a white solid is obtained by filtration, which is washed with ethyl acetate several times and then dried under vacuum at room temperature to obtain a white solid (see Figure 5).

[0139] 3. Synthesis of Compound 3

[0140] (1) Dissolve Boc-Lys(Boc)-OH, EDC, HOBT and compound 2 in dry dichloromethane.

[0141] (2) In an ice-water bath, slowly add DIPEA or triethylamine dropwise. The molar ratio of the raw materials is compound 2: Boc-Lys(Boc)-OH: EDC: HOBT: DIPEA or triethylamine = 1:2:2:2:2. Remove the ice-water bath and continue stirring the reaction for 24 h.

[0142] (3) After the reaction is completed, the mixture is washed with dilute hydrochloric acid, dilute sodium hydroxide, and saturated brine, respectively. The organic phase is separated, dried over anhydrous magnesium sulfate, filtered, and spin-dried. The crude product is separated and purified by silica gel column chromatography to obtain a white solid (see Figure 6).

[0143] 4. Synthesis of Compound 5

[0144] (1) Compound 1 or 3 was dissolved in a mixed solvent of methanol / water = 4:1.

[0145] (2) Under ice-water bath conditions, slowly add lithium hydroxide or sodium hydroxide or potassium hydroxide, with the molar ratio of the raw materials being compound 1 or 3: lithium hydroxide or sodium hydroxide or potassium hydroxide = 1:2.

[0146] (3) Remove the ice-water bath and continue the reaction for 12 h. Then, add dilute hydrochloric acid to the reaction solution to adjust the pH to 5.

[0147] (4) After removing the methanol in the system under reduced pressure, the product was extracted with ethyl acetate three times, and the organic phases were combined and dried by spin drying to finally obtain a white product (see Figures 7 and 8).

[0148] 5. Synthesis of Compounds 6-8 (Ma)

[0149] (1) Compound 4 or 5 or Boc-Glc-OH and EDC were dissolved in dry dichloromethane, and pentafluorophenol was slowly added in an ice-water bath.

[0150] (2) The molar ratio of the raw materials is compound 4 or 5 or Boc-Glc-OH:EDC:pentafluorophenol = 1:2:2. After removing the ice-water bath, the reaction is stirred at room temperature for 12 h.

[0151] (3) After the reaction was completed, the excess solvent was removed by distillation under reduced pressure, and the crude product was separated by silica gel column chromatography to obtain a white solid (see Figures 9, 10, and 11).

[0152] 6. Synthesis of Compound 9

[0153] (1) Dissolve Boc-Lys(Boc)-OH, EDC, HOBT, and propargylamine in dry dichloromethane. Slowly add DIPEA or triethylamine dropwise in an ice-water bath.

[0154] (2) The molar ratio of the raw materials is Boc-Lys(Boc)-OH:EDC:HOBT:propargylamine:DIPEA or triethylamine = 1:1:1:1:1. After removing the ice-water bath, the reaction is continued for 24 hours.

[0155] (3) After the reaction, the system was washed with dilute hydrochloric acid and dilute sodium hydroxide solution, and the organic phase was separated. The crude product was separated and purified by silica gel column chromatography to obtain the product (see Figure 12).

[0156] 7. Synthesis of Compound 10 (Mb)

[0157] (1) Compound 9 was cooled in an ice-water bath, and then an excess of hydrogen chloride saturated ethyl acetate solution was slowly added dropwise to dissolve it. The ice-water bath was removed and the reaction was continued for 0.5 to 6 hours.

[0158] (2) After the reaction is completed, the white solid is filtered and washed with ethyl acetate several times and then dried in a vacuum oven to obtain the product (see Figure 13).

[0159] 8. Synthesis of Compound 11 (Mc):

[0160] (1) Dissolve Boc-aminoacetic acid and EDC in dry dichloromethane. Slowly add pentafluorophenol in an ice-water bath. The molar ratio of the raw materials is Boc-aminoacetic acid: EDC: pentafluorophenol = 1:1:1.

[0161] (2) After removing the ice-water bath, continue stirring the reaction for 24 h. After the reaction is complete, the precipitate is filtered, washed several times with dichloromethane, and vacuum-dried to obtain a white product (see Figure 14).

[0162] 1.4 Preparation and Characterization of P0, P1, P2, Component A (including Component A with Sugar Molecules Changed to Glucose) (P0-Man(Glc), P1-Man(Glc), P2-Man(Glc)), and Component B (P0-Bob, P1-Bob, P2-Bob) (Synthesis Route See Figure 15)

[0163] 1.4.1 Synthesis of Compounds 12, 13, and 14 (P0, P1, and P2)

[0164] (1) Compound 6, 7, or 8 and Compound 10 were slowly dissolved in anhydrous DMSO or DMF. DIPEA or triethylamine was slowly added dropwise in an ice-water bath.

[0165] (2) The molar ratio of the raw materials is Compound 6, 7, or 8: Compound 10: DIPEA or triethylamine = 1:1:2. After removing the ice-water bath, continue stirring the reaction for 48 hours.

[0166] (3) After the reaction is complete, add methanol or ethanol to dilute the solution under ice-water bath conditions, then precipitate the product in ether and centrifuge. Repeat this step three times, and finally filter the precipitate and vacuum dry to obtain the product (see Figure 16).

[0167] 1.4.2 Synthesis of Compounds 15, 16, and 17

[0168] (1) Compound 12, 13, or 14 was dissolved in excess trifluoroacetic acid in an ice-water bath. After removing the ice-water bath, stirring was continued for 4.5 h.

[0169] (2) After the reaction, the solution was diluted with methanol and the product was precipitated in diethyl ether. The product was filtered out, washed with diethyl ether several times and then dried in vacuum to obtain a white product (see Figures 17, 18, and 19).

[0170] 1.4.3 Synthesis of Compounds 19 and 20 (P0-Bob, P1-Bob, P2-Bob)

[0171] (1) Add compound 15, 16, or 17, phenylboronic acid half ester, and sodium borohydride into a flask and evacuate the mixture.

[0172] (2) The molar ratio of the raw materials is compound 15, 16, or 17: phenylboronic acid half ester: sodium borohydride = 1:1:4.

[0173] (3) Then, in an ice-water bath, methanol was slowly added dropwise. The ice-water bath was then removed and the reaction was continued with stirring for 24 h.

[0174] (4) After the reaction, the product was dialyzed in deionized water using a dialysis bag with a molecular weight cutoff of less than 2000 and freeze-dried to obtain a white solid (see Figures 20, 21, and 22).

[0175] 1.4.4 Synthesis of Compounds 21, 22, and 23

[0176] (1) Compound 15, 16, or 17 and compound 11 (Mc) were dissolved in anhydrous DMSO or DMF.

[0177] (2) In an ice-water bath, slowly add DIPEA or triethylamine dropwise. The ratio of the starting materials is compound 15, 16, or 17: compound 11: DIPEA or triethylamine = 1:1:2.

[0178] (3) Remove the ice-water bath and continue stirring the reaction for 24 h.

[0179] (4) After the reaction is complete, dilute the solution with methanol in an ice-water bath and precipitate the product with ether. Repeat this process three times. Finally, filter the precipitate and dry the white solid in a vacuum oven at room temperature.

[0180] (5) Then, dissolve the solid in excess trifluoroacetic acid in an ice-water bath. Remove the ice-water bath and continue stirring for 4.5 h.

[0181] (6) After the reaction, the product was diluted with methanol in an ice-water bath and precipitated with ether to separate the product. After filtration, it was washed with ether several times and dried under vacuum at room temperature to obtain a white solid (see Figures 23, 24, and 25).

[0182] 1.4.5 Synthesis of Compounds 24, 25, and 26 (P0-Man(Glc), P1-Man(Glc), and P2-Man(Glc) (Synthesis Route See Figure 30)

[0183] (1) Compound 21, 22, or 23, mannose (glucose), and aniline (10-100 μL) were dissolved in an acetic acid / sodium acetate buffer solution at pH 4.2 and stirred at 37° C. for 6-24 h.

[0184] (2) The molar ratio of the raw materials is compound 21, 22, or 23: mannose (glucose) = 1:1.

[0185] (3) Dialysis was performed using a dialysis bag with a molecular weight cut-off of less than 2000, and freeze-dried to obtain white solids (26, 27, 28, 29, 30). 1.4.6 Synthesis of compound 27 (P0-Bob-1)

[0186] (1) Compound 15 (0.155 g, 0.53 mmol) and Bob-1 (0.199 g, 0.58 mmol) were dissolved in 1.5 mL of anhydrous DMF. After cooling in an ice-water bath, DIPEA (0.3 mL, 1.63 mmol) was slowly added dropwise to the system. The ice-water bath was removed and stirred for 6 h. After the reaction was completed, the reaction system was diluted with 5 mL of methanol and then precipitated with 250 mL of diethyl ether. This step was repeated three times to fully remove DIPEA. The product was finally dried in vacuum to obtain a white solid (0.197 g, 82%).

[0187] 1.4.7 Synthesis of Compound 28 (P1-Bob-1)

[0188] (1) Compound 16 (0.165 g, 0.39 mmol) and Bob-1 (0.295 g, 0.86 mmol) were dissolved in 1.5 mL of anhydrous DMSO. After cooling in an ice-water bath, DIPEA (0.3 mL, 1.63 mmol) was slowly added dropwise to the system. The ice-water bath was removed and stirred for 6 h.

[0189] (2) After the reaction is complete, dilute the reaction system with 5 mL of methanol and then precipitate with 250 mL of ether. Repeat this step 2 to 3 times to fully remove DIPEA.

[0190] (3) The product was finally dried under vacuum to obtain a white solid (0.219 g, 76%).

[0191] 1.4.8 Synthesis of Compound 29 (P2-Bob-1)

[0192] (1) Compound 17 (0.130 g, 0.19 mmol) and Bob-1 (0.297 g, 0.86 mmol) were dissolved in 1.5 mL of anhydrous DMF. After cooling in an ice-water bath, DIPEA (0.3 mL, 1.63 mmol) was slowly added dropwise to the system. The ice-water bath was removed and stirred for 6 h.

[0193] (2) After the reaction, the reaction system was diluted with 5 mL of methanol and then precipitated with 250 mL of ether. This step was repeated three times to fully remove DIPEA. Finally, the product was vacuum dried to obtain a white solid (0.182 g, 72%) (see Figure 31). 1.4.9 Synthesis of Compound 30 (P2-Man-RGD)

[0194] (1) Compound 26, N3-GRGDSP peptide, DMF, cuprous bromide, and PMDETA were added to a flask and deoxygenated.

[0195] (2) Heat to 50 degrees Celsius and continue stirring to react for 24 hours.

[0196] (3) After the reaction, the product was dialyzed in deionized water using a dialysis bag with a molecular weight cut-off of less than 2000 and freeze-dried to obtain a white solid.

[0197] The product P2-Man-RGD was a white solid ( FIG. 32 ).

[0198] 1.5 Preparation and Characterization of G0, G1, and G2 with Different Branching Patterns (See Figure 33)

[0199] 1.5.1 Synthesis of G1.0

[0200] (1) The polymer backbone G0 and the monomer methyl acrylate MA were dissolved in anhydrous methanol at room temperature, and then the methanol solution of G0 was slowly dripped into the methanol solution of MA. The molar ratio of the raw materials was G0 (n-NH2): MA = 1:30.

[0201] (2) After the addition is complete, the mixture is stirred at room temperature for 48 hours. After the reaction is complete, the unreacted MA and the solvent methanol are removed by spin drying to obtain product G0.5.

[0202] (3) G0.5 and the reaction raw material ethylenediamine were then dissolved in anhydrous methanol at room temperature, and then the methanol solution of G0.5 was slowly dripped into the methanol solution of ethylenediamine. The molar ratio of the raw materials was G0.5 (n-COOR): ethylenediamine = 1:50.

[0203] (4) After the addition is complete, the mixture is stirred at room temperature for 48 hours. After the reaction is complete, the unreacted ethylenediamine and the solvent methanol are removed by spin drying to obtain product G1.0 (see Figures 34 and 35).

[0204] 1.5.2 Synthesis of G2.0

[0205] (1) Dissolve the polymer G1.0 and the monomer methyl acrylate (MA) in anhydrous methanol at room temperature. Then slowly drip the G0 methanol solution into the MA methanol solution. The molar ratio of the raw materials is G1.0 (n-NH2):MA = 1:50. After the dripping is complete, stir and react at room temperature for 48 hours.

[0206] (2) After the reaction, the unreacted MA and the solvent methanol were dried by spin drying to obtain the product G1.5 (see Figure 36). Subsequently, G1.5 and the reaction raw material ethylenediamine were dissolved in anhydrous methanol at room temperature. The methanol solution of G1.5 was then slowly added dropwise to the methanol solution of ethylenediamine. The molar ratio of the raw materials was G1.5 (n-COOR): ethylenediamine = 1:50.

[0207] (3) After the addition is complete, the mixture is stirred at room temperature for 48 hours. After the reaction is complete, the unreacted ethylenediamine and the solvent methanol are removed by spin drying to obtain product G2.0 (see Figure 37).

[0208] Example 2 Hydrogel Preparation and Properties

[0209] 2.1 Gel preparation method

[0210] First, polymer solutions of component A (Pn-Man, Pn-Glc (n=0, 1, 2)) and component B (Pn-Bob (n=0, 1, 2)) were prepared: the solid content of solutions A and B was controlled to be 12wt% to 20wt%. Hyperbranched glycopeptide polymers (P0-Man, P1-Man, P2-Man) were dissolved in pure water to obtain component A solution, and hyperbranched phenylboronic acid half ester polypeptides (P0-Bob, P1-Bob, P2-Bob) were dissolved in pure water and the pH was adjusted to 8 to obtain component B solution. The donor / acceptor (sugar unit / phenylboronic acid unit) ratio in the solutions of components A and B was adjusted to 2:1. After the two solutions were completely dissolved in the solvent, the two solutions were mixed evenly and allowed to stand to obtain a series of hyperbranched polypeptide hydrogel materials.

[0211] 2.2 Gel performance test

[0212] 2.1.1 Hydrogel adhesion stability test

[0213] Pigskin was trimmed into 1 cm x 1 cm sections and fixed to a 1 cm x 4 cm acrylic plate using quick-drying adhesive. A total volume of 20 μL of hydrogel was applied between the two pieces of pigskin. After waiting 10 minutes for the gel to fully solidify, the ends of the specimen were secured with clamps. The specimen was pulled using a ZP50 tensile testing machine at a speed of 2 mm / min until complete separation. The maximum tissue adhesion strength achieved before complete separation was recorded.

[0214] The experimental results of the binding ability of the two components showed that increasing the degree of branching can effectively increase the number of terminal functionalized sites, thereby increasing the number of bindings, and ultimately further increasing the molar binding enthalpy of the two, that is, the gel stability is better (see Figure 38a, Figure 38a); P1-Glc+P1-Bob and P1-Man+P1-Bob have almost the same binding exotherm, indicating that their binding abilities are very close (see Figure 38c).

[0215] 2.1.2 Shear test

[0216] The gel can provide effective adhesion within the ratio range of 1:2 and 2:1, has good tissue adhesion and shear resistance, and is suitable for applications in skin wound repair and conjunctival transplantation (see Figure 39).

[0217] The rotational rheometer test shows that under different combinations of A and B, the gel strength can be precisely controlled in the range of 10Pa to 100,000Pa (see Figures 40 and 41). The rotational rheometer test shows that, when neutral, the mechanical strength of the gels formed by P1-Glc+P1-Bob and P1-Man+P1-Bob are almost the same (see Figure 42).

[0218] 2.1.3 Torsion test - hydrogel adhesion stability test on pig skin in vitro under external force

[0219] First, the gel was stained with Macaus blue for easy observation. A total of 20 μL of the two-component polymer solution, A and B, was dropped onto pig skin. After waiting 10 minutes for the solution to completely solidify in situ to form a hydrogel, the pig skin was subjected to various stresses, including twisting, folding, and stretching. The gel was observed for adhesion stability under stress and for any signs of detachment or cracking, and photographed.

[0220] The gel applied to the pigskin surface remained stably attached to the surface under various deformation states without falling off or breaking. The gel has good adhesion stability (see Figure 43).

[0221] 2.1.4 Adhesion stability - underwater test Gel adhesion stability test

[0222] The hydrogel was stained with Macaus blue to facilitate subsequent observation. After the polymer solution was dropped onto pigskin to form a hydrogel for in-situ polymerization, the hydrogel was then immersed in a PBS buffer solution (pH 7.2), with the solution changed daily until the seventh day. After immersion, the sample was removed and observed for its ability to stably adhere to the pigskin surface after prolonged immersion in water, as well as its adhesion stability under running water.

[0223] The gel can still stably adhere to the tissue surface after being soaked in water for a long time. The gel has good adhesion stability (see Figure 44).

[0224] 2.1.5 Light transmittance test

[0225] The transmittance test results preliminarily show that the visible light transmittance of the 0+0 formula is close to that of carbomer and better than that of the 1+1 formula. However, in actual imaging, the flatness of 0+0 is not as good as that of carbomer, and it is easy to produce small wrinkles, so the imaging transmittance is slightly inferior to that of carbomer gel (see Figure 45); among them, the 0+0 formula is a formula of component A (Pn-Man, Pn-Glc (n=0)) and component B (Pn-Bob (n=0)); the 1+1 formula is a formula of component A (Pn-Man, Pn-Glc (n=1)) and component B (Pn-Bob (n=1)); the 0+1 formula is a formula of component A (Pn-Man, Pn-Glc (n=1)) and component B (Pn-Bob (n=0).

[0226] 2.1.6 In vitro degradation experiment

[0227] The results showed that the gel had certain stability in a wet environment and medium- to long-term degradation (see Figure 46).

[0228] 2.1.7 Gel for Autologous Conjunctival Transplantation Testing

[0229] Symmetrical 1 cm x 1 cm conjunctival defects were created in pig eyes. In the experimental group, 12 μL of hydrogel was injected into the conjunctival defect, followed by adhesion of a conjunctival flap after 1 minute; in the control group, the flap was left to naturally cover. Images were taken at 0, 10, 20, 30, and 40 minutes, and the displacement area of ​​the conjunctival flap was analyzed using Image J. A further conjunctival defect identical to the above was created. The experimental group adhered the conjunctival flap with the hydrogel, while the control group allowed natural coverage. After transplantation, the conjunctival graft site was continuously irrigated with water, and the grafted conjunctival flap was observed for detachment to assess adhesion stability.

[0230] The 0+0 and 1+1 gel formulas have good adhesion and can effectively fix the conjunctiva and sclera; the flushing experiment shows that the 0+0 and 1+1 gel formulas have good adhesion, which can make the transplanted conjunctival tissue adhere to the original position and not be washed away by water (see Figure 47).

[0231] 2.1.8 Biomechanical testing of hydrogel-sealed corneal incisions in porcine eyes in vitro

[0232] According to the corneal incision closure method, pig eyes were randomly divided into 4 groups: suture group, Gel0+0 (component A (Pn-Man, Pn-Glc (n=0)) and component B (Pn-Bob (n=0))), Gel1+1 (component A (Pn-Man, Pn-Glc (n=1)) and component B (Pn-Bob (n=1)), fibrin group. The corneal flap was removed using a corneal trephine under an operating microscope and placed in an artificial anterior chamber. The water inlet end of the artificial anterior chamber was connected to a microinjection syringe. The injection pump was connected to a differential pressure gauge at the outlet. After the artificial anterior chamber was filled to 21 mmHg (normal intraocular pressure), a 2.8 mm linear incision was made near the limbus using a corneal primary incision knife. The corneal incision was sealed using either the experimental group hydrogel, commercial fibrin glue, or sutures. After the procedure, the microinjection pump was turned on to inject fluid into the artificial anterior chamber at a pumping rate of 10 mm / h. The differential pressure gauge reading was recorded by video. When the peak value appeared, the incision was observed for fluid leakage and the reading was recorded.

[0233] The results showed that both the 0+0 and 1+1 formulations exhibited less leakage than the suture group and commercial Fibrin, demonstrated better sealing effects, and could withstand intraocular pressures exceeding 200 mmHg, making them suitable for corneal wound closure (see Figure 48).

[0234] Example 3 Biological experiments of gel

[0235] 3.1 Biosafety Testing

[0236] SD rats were intradermally injected with 100 μl of P1Bob + 100 μl of P1Man prepared in Example 1, and samples were taken for pathological sections 2 weeks, 1 month, 2 months, and 3 months after the injection.

[0237] The results showed that there was no significant difference between the experimental group and normal skin, and no obvious inflammatory reaction was caused, proving that the gel has good biocompatibility (see Figure 49).

[0238] The results of the cell safety test showed that the material did not affect cell activity and normal cell proliferation (see Figure 50).

[0239] 3.2 Rabbit autologous conjunctival transplantation

[0240] Suture, 0+0 (A component (Pn-Man, Pn-Glc (n=0)) and B component (Pn-Bob (n=0)), 1+1 (A component (Pn-Man, Pn-Glc (n=1)) and B component (Pn-Bob (n=1)), and commercial materials were compared respectively.

[0241] Preliminary results showed that the gel group had a transplantation effect. The tissue at the transplant site in the gel group showed a tendency to repair. Conjunctival morphology of stratified squamous epithelium and a small amount of goblet cells were visible. The epithelial regeneration effect was good, equivalent to that of the suture group and better than that of commercialized Fibrin (see Figure 51).

[0242] Example 4 Animal Model of Corneal Injury Repair

[0243] A 2.8 mm single-plane linear incision was made near the limbus of the rabbit eyes, and then closure was performed with nylon sutures or gel according to the groups.

[0244] The results of intraocular pressure monitoring showed that the intraocular pressure of all groups basically returned to normal three days after the incision was glued, indicating that the treatment methods of all groups could effectively seal the incision and prevent anterior chamber leakage.

[0245] Anterior segment optical coherence tomography showed that the Gel 1+1 (component A (Pn-Man, Pn-Glc (n=1)) and component B (Pn-Bob (n=1)) groups had a more significant decrease in internal and external incision thickness at 3-7 days compared with the other groups, which can promote early healing of corneal tissue and disappearance of edema earlier.

[0246] The slice results showed that the Gel 1+1 group had the best repair effect, the tissue structure at the incision was continuous, and the collagen fibers in the corneal stroma were arranged regularly and had a normal structure.

[0247] In general, Gel 1+1 can effectively seal the corneal incision, facilitate early corneal tissue repair and edema reduction, and contribute to the correct morphological distribution of the cornea after surgery, with the best repair effect (see Figure 52).

[0248] Example 5 Bone defect repair animal model

[0249] A 3mm diameter and 3mm deep bone defect was created in the left femoral condyle of SD rats using a dental drill. Four groups of rats received the following implants to repair the bone defect:

[0250] (1) Pathology group: debridement was performed only at the defect site;

[0251] (2) Gel 1+1 (Component A (Pn-Man, Pn-Glc (n=1)) and Component B (Pn-Bob (n=1)) group: 40 μl of the 1+1 gel complex was implanted in the defect;

[0252] (3) Gel 2+2 (Component A (Pn-Man, Pn-Glc (n=2)) and Component B (Pn-Bob (n=2)) groups: 40 μl of the 2+2 gel complex was implanted in the defect;

[0253] (4) 2+2 RGD group ((Component A (Pn-Man-RGD (n=2)) and Component B (Pn-Bob (n=2))): 40ul of the 2+2 RGD gel complex was implanted in the defect.

[0254] At week 4, the left femoral condyle of the rats was examined using Micro-CT.

[0255] CT reconstruction results showed (see Figure 53) that GEL2+2 and GEL2+2 RGD treatment groups significantly promoted bone tissue defect repair compared with the blank group and GEL1+1 group.

[0256] The above description is a general description of the present invention. Variations in form and substitutions of equivalents may be made as appropriate. Although specific terminology is employed herein, such terminology is intended to be descriptive, not restrictive. Those skilled in the art may make various changes or modifications to the present invention, and such equivalents are intended to fall within the scope of the appended claims.

Claims

1. A hydrogel, the hydrogel is composed of two polymers A and B, the main chains of the two polymers A and B are composed of amide bonds formed by lysine and glutamic acid through polycondensation, and the value of n is 8-100; the two polymers A and B are polymers with arbitrary branching generations; the structural formulas of the two polymers A and B are shown in a and b respectively: The R a Selected from (c) or (d), the structural formula is as shown in (c) and (d): or In the formula, Ra1 is the alkyne terminal on the main chain of the two polymers A and B connected to the exogenous functional molecules; R b It is the branched structure of the amino terminal of the side chain on the two polymer backbones A and B; R c R b Terminally linked monosaccharide or oligosaccharide; R d R b Any phenylboronic acid structure compound connected to the end.

2. The hydrogel according to claim 1, characterized in that The functional molecules are selected from one or more of peptides, fluorescent groups, antibacterial molecules, growth factors, functional oligosaccharides or polysaccharides.

3. The hydrogel according to claim 1, characterized in that The monosaccharide connected to the polymer A can be selected from one or more of mannose, glucose, galactose, fructose, xylose, fucose, gulose, idose, lyxose, mannuronic acid, glucuronic acid, galacturonic acid, guluronic acid and the like and their derivatives, and the oligosaccharide connected to the polymer A can be one or more of any oligosaccharide.

4. The hydrogel according to claim 1, characterized in that Any phenylboronic acid structure connected to the polymer B includes one or more of phenylboronic acid half ester and carbonyl phenylboronic acid half ester.

5. The hydrogel according to claim 1, characterized in that The ratio of polymer A to polymer B in the hydrogel is 1-2:2-1.

6. Use of the hydrogel according to claims 1 to 5 in the preparation of skin wound materials.

7. Use of the hydrogel according to claims 1 to 5 in preparing a suture required for conjunctival transplantation or corneal transplantation.

8. A medical preparation comprising the hydrogel according to claims 1-5.

9. Use of the hydrogel according to claims 1 to 5 in preparing bone repair materials.

10. A method for preparing the hydrogel according to claims 1-5: S1. Preparation of intermediate 1: The raw materials Boc-Lys(Boc)-OH and H-Glu(OMe)-OMe·HCl underwent condensation reaction under the action of reagents EDC, HOBT and DIPEA to obtain intermediate 1; S2. Preparation of intermediate product 2: The intermediate product 1 undergoes a deprotection reaction under the action of a reagent, hydrogen chloride ethyl acetate solution, to obtain the intermediate product 2; S3. Preparation of intermediate product 3: Intermediate product 2 and raw material Boc-Lys(Boc)-OH undergo condensation reaction under the action of reagents EDC, HOBT and DIPEA to obtain intermediate product 3; S4. Preparation of intermediate products 4 and 5: The intermediate product 1 or 3 undergoes a deprotection reaction under the action of lithium hydroxide reagent to obtain the intermediate product 4 or 5 respectively; S5. Preparation of intermediate products 6, 7, and 8: The raw material Boc-Glu-OH or the intermediate product 4 or 5 and the raw material pentafluorophenol undergo condensation reaction under the action of the reagent EDC to obtain the intermediate products 6, 7, and 8 respectively; S6. Preparation of intermediate product 9: The raw material Boc-Lys(Boc)-OH and propargylamine reacted with reagents EDC, HOBT and DIPEA to obtain the intermediate 9; S7. Preparation of intermediate product 10 (Mb): The intermediate product 9 undergoes a deprotection reaction under the action of a reagent, hydrogen chloride ethyl acetate solution, to obtain the intermediate product 10 (Mb); S8. Preparation of intermediate product 11 (Mc): The raw materials Boc-aminooxyacetic acid and pentafluorophenol underwent condensation reaction under the action of the reagent EDC to obtain the intermediate product 11 (Mc); S9. Preparation of intermediates 12, 13, 14: Intermediate product 6 or 7 or 8 and intermediate product 10 (Mb) undergo polymerization reaction under the action of reagent DIPEA to obtain intermediate products 12, 13, and 14; S10. Preparation of intermediates 15, 16, and 17: The intermediate product 12 or 13 or 14 undergoes a deprotection reaction under the action of trifluoroacetic acid to obtain intermediate products 15, 16, and 17, respectively; Preparation of S11.B component (products 18, 19, 20): The intermediate product 15 or 16 or 17 reacts with the raw material Bob under the action of the reagent sodium borohydride to obtain the product B component (product 18, 19, 20) respectively; S12. Preparation of intermediates 21, 22, and 23: Intermediate product 15 or 16 or 17 and intermediate product 11 (Mc) undergo condensation reaction under the action of reagent DIPEA, and further undergo deprotection reaction under the action of reagent trifluoroacetic acid to obtain intermediate products 21, 22, and 23, respectively; Preparation of S13.A component (products 24, 25, 26): The intermediate product 21 or 22 or 23 and the raw material monosaccharide or oligosaccharide undergo condensation reaction under the action of the reagent aniline to obtain product A component (product 24, 25, 26) respectively; S14. Synthesis of hydrogel: The solid content of component A or component B is prepared at 12wt% to 20wt%.

Images