CN114763391A

CN114763391A - Compound or polymer containing phenolic adhesion group and method for functionalizing hydrogel

Info

Publication number: CN114763391A
Application number: CN202110043722.5A
Authority: CN
Inventors: 刘润辉; 张东辉; 刘晶晶
Original assignee: East China University of Science and Technology
Current assignee: East China University of Science and Technology
Priority date: 2021-01-13
Filing date: 2021-01-13
Publication date: 2022-07-19
Anticipated expiration: 2041-01-13
Also published as: CN114763391B

Abstract

The invention discloses a compound or polymer containing phenolic adhesive groups and a method for functionalizing hydrogel, and the method comprises the following stepsThe phenolic adhesion group consists of repeating units shown in general formulas I and II which are arranged in a random, block or alternating mode, R is a positive charge side group, and the total number x of the repeating units shown in the general formula I is an integer of 0-200; the total number y of the repeating units represented by the general formula II is an integer of 1 to 200. By adopting the polymer, the hydrogel can be functionalized in a simple manner, namely, the surface and the interior of hydrogels of different types can be modified in one step, and the functional hydrogel of variable types can be prepared, and the hydrogel can be functionalized without specific functionalization in the whole process.

Description

Compound or polymer containing phenolic adhesive group and method for functionalizing hydrogel

Technical Field

The present invention relates to a compound or polymer containing phenolic adhesive groups and a method for functionalizing a hydrogel, i.e., a method for preparing a functional hydrogel, using the same.

Background

The hydrogel has the characteristics of high hydration and inertia three-dimensional network, and is widely applied to the field of biological materials. For different biomaterials requiring hydrogels to perform different functions, different functional molecules need to be introduced into the hydrogel. Currently, these functionalized hydrogels are prepared by two routes: first, a hydrogel precursor is chemically modified with functional molecules and then gelated (adv. drug delivery. rev.64,223-236 (2012)); alternatively, a hydrogel with specific reactive sites is prepared and then functional molecules are modified to the reactive sites of the hydrogel network (Chinese J. Polym. Sci.35,1181-1193 (2017)). However, the functional molecules and functional groups of both methods are specific and require separate synthesis of compounds of functional or reactive groups for each type of hydrogel, steps that are cumbersome and very inflexible. Therefore, there is an urgent need to establish a general strategy for the preparation of functional hydrogels of varying types.

Disclosure of Invention

The aim of the present invention is to establish a general strategy for the preparation of functional hydrogels of variable type.

In a first aspect of the invention, there is provided a compound or polymer containing a phenolic adhesion group consisting of repeating units represented by the general formulae I and II:

in the formula, R is a positive charge side group and is a side group containing primary amine, secondary amine or tertiary amine;

the arrangement form of the repeating units represented by the general formulas I and II is random, block or alternate, wherein the total number x of the repeating units represented by the general formula I is an integer of 0-200; the total number y of the repeating units represented by the general formula II is an integer of 1 to 200.

In another preferred embodiment, x is an integer of 0 to 100, preferably 0 to 50 or 1 to 30.

In another preferred embodiment, y is an integer of 1 to 100, preferably 1 to 50 or 5 to 30

In another preferred embodiment, the amino acids corresponding to the repeating units represented by formula I include, but are not limited to, lysine, ornithine, arginine.

In another preferred embodiment, R is C1-C18 alkyl substituted by amino, C1-C18 alkyl substituted by mono-C1-C18 alkylamino, C1-C18 alkyl substituted by di-C1-C18 alkylamino.

In another preferred embodiment, R is C1-C10 alkyl substituted by amino, C1-C10 alkyl substituted by mono (C1-C10 alkyl) amino, C1-C10 alkyl substituted by di (C1-C10 alkyl) amino.

In another preferred embodiment, R is C1-C6 alkyl substituted by amino, C1-C6 alkyl substituted by mono (C1-C6 alkyl) amino, C1-C6 alkyl substituted by di (C1-C6 alkyl) amino.

In another preferred embodiment, the phenolic adhesion groups are:

in another preferred embodiment, the anion of the above structure is selected from: cl^-、Br^-、CF₃COO^-、H₂PO₄ ^-、HPO₄ ^2-、PO₄ ^3-。

In another preferred embodiment, the structure of the compound or polymer is represented by formula V:

wherein R, x and y are as defined above;

-/- (-in the arrangement in random, block or alternate form;

R₁h, C1-C18 alkyl, amino-substituted C1-C18 alkyl, hydroxyl, amino, carboxyl, aldehyde, C2-C18 alkenyl, sulfhydryl, azide, C2-C18 alkynyl, ortho-dithiopyridyl (OPSS); or a group containing maleimide, biotin, adamantane, cyclodextrin; or a polymer chain selected from amino acid polymer chains and carbon chain polymer chains;

R₂h, C1-C18 alkyl, amino-substituted C1-C18 alkyl, hydroxyl, amino, carboxyl, aldehyde, C2-C18 alkenyl, sulfhydryl, azide, C2-C18 alkynyl, ortho-dithiopyridyl (OPSS); or a group containing maleimide, biotin, adamantane, cyclodextrin; or polymer chain selected from amino acid polymer chain and carbon chain polymer chain.

In another preferred embodiment, R₁Is C1-C10 alkyl substituted by amino, C1-C10 alkyl substituted by mono (C1-C10 alkyl) amino, C1-C10 alkyl substituted by di (C1-C10 alkyl) amino; more preferably, R is C1-C6 alkyl substituted by amino, C1-C6 alkyl substituted by mono (C1-C6 alkyl) amino, C1-C6 alkyl substituted by di (C1-C6 alkyl) amino.

In another preferred embodiment, R₂Is C1-C10 alkyl substituted by amino, C1-C10 alkyl substituted by mono (C1-C10 alkyl) amino, C1-C10 alkyl substituted by di (C1-C10 alkyl) amino; more preferably, R is amino-substituted C1-C6 alkyl, mono (C1-C6 alkyl) amino-substituted C1-C6 alkyl, di (C1-C6 alkyl) amino-substituted C1-C6 alkyl

In another preferred embodiment, the compound or polymer has the structure

In the formula, R₂The definition of (c) is as described above.

At another placeIn a preferred embodiment, the anion of the above structure is selected from: cl^-、Br^-、CF₃COO^-、H₂PO₄ ^-、HPO₄ ^2-、PO₄ ^3-。

In another preferred embodiment, the polymer is selected from the group consisting of:

in each formula, each n is independently an integer of 1 to 1000.

In another preferred embodiment, the beta-amino acid side groups of Pol-7 and Pol-8 are racemates.

In a second aspect of the present invention, there is provided a process for the preparation of a polymer containing phenolic adhesion groups as described in the first aspect, said process being obtained by initiating a polymerisation reaction by means of an initiator selected from the group consisting of:

in a third aspect of the invention, there is provided an initiator selected from the group consisting of:

in a fourth aspect of the invention, there is provided the use of a polymer comprising phenolic adhesive groups according to the first aspect for modifying a hydrogel or for adhering to a material surface.

In another preferred embodiment, the hydrogel is selected from the group consisting of: polyethylene glycol (PEG) hydrogel, polyvinyl alcohol (PVA) hydrogel, Polyhydroxyethylmethacrylate (PHEMA) hydrogel, Polysulfonobetaine (PSBMA) hydrogel, polycarboxybetaine (pcbmma) hydrogel, alginic Acid (ALG) hydrogel, Hyaluronic Acid (HA) hydrogel, dextran hydrogel, chitosan hydrogel.

In another preferred example, the material surface is a metal surface, a metal oxide surface, an inorganic non-metal surface, or a polymer material surface.

In another preferred embodiment, the metal is selected from: gold, silver, copper, iron, stainless steel, aluminum, titanium.

In another preferred embodiment, the metal oxide is selected from: iron oxide, titanium oxide, zirconium oxide.

In another preferred embodiment, the inorganic nonmetal is selected from the group consisting of: glass, silicon oxide, mica.

In another preferred embodiment, the polymer is selected from: polyethylene, polypropylene, polymethyl methacrylate, polyester, polystyrene, polyether ether ketone, polytetrafluoroethylene, polyurethane.

In a fifth aspect of the present invention, there is provided a functional hydrogel comprising a hydrogel matrix modified at the surface and/or interior with the polymer comprising phenolic adhesive groups of the first aspect.

In another preferred embodiment, the hydrogel matrix is selected from the group consisting of: polyethylene glycol (PEG) hydrogel, polyvinyl alcohol (PVA) hydrogel, Polyhydroxyethylmethacrylate (PHEMA) hydrogel, Polysulfonobetaine (PSBMA) hydrogel, polycarboxybetaine (pcbmma) hydrogel, alginic Acid (ALG) hydrogel, Hyaluronic Acid (HA) hydrogel, dextran hydrogel, chitosan hydrogel.

In a sixth aspect of the present invention, there is provided a method for preparing the functional hydrogel according to the fifth aspect, comprising the step of immersing a hydrogel in a solution of the polymer containing phenolic adhesion groups to obtain the functional hydrogel.

In another preferred embodiment, the concentration of the solution of the compound containing phenolic adhesion groups is 0.01-10 mg/mL.

In a seventh aspect of the present invention, there is provided a use of the functional hydrogel according to the fifth aspect for preparing an antibacterial material or preparing a tissue engineering scaffold.

In an eighth aspect of the present invention, there is provided a method for general surface modification and functionalization of a specific end group, wherein the specific end group has the following structure:

the compound or polymer containing the terminal group can adhere to various surfaces, including metal surfaces (gold, silver, copper, iron, stainless steel, aluminum, titanium, etc.), metal oxide surfaces (iron oxide, titanium oxide, zirconium oxide, etc.), inorganic nonmetal surfaces (glass, silicon oxide, mica, etc.), polymer material surfaces (polyethylene, polypropylene, polymethyl methacrylate, polyester, polystyrene, polyether ether ketone, polytetrafluoroethylene, polyurethane, etc.), and the like.

According to the invention, compounds containing phenolic adhesion groups are modified to the surfaces and the interiors of different types of hydrogels in one step, and the whole process can realize the functionalization of the hydrogel without specific functionalization, so that multiple functions are provided for the hydrogel, such as the functions of antibiosis, cell adhesion, wound healing and the like of the hydrogel.

It is to be understood that within the scope of the present invention, the above-described features of the present invention and those specifically described below (e.g., in the examples) may be combined with each other to form new or preferred embodiments. Each feature disclosed in this specification may be replaced by an alternative feature serving the same, equivalent, or similar purpose. Not to be reiterated herein, but to the extent of space.

Detailed Description

The present inventors have extensively and intensively studied to synthesize Dba-containing compoundYKYCompounds of the group which have been found to functionalize water in one stepThe functionalized hydrogel has the functions of antibiosis, cell adhesion, wound healing and the like on the surface and in the gel. On the basis of this, the present invention has been completed.

DbaYKYSynthesis of intermediates

Synthesis of Compound 1

The starting material Fmoc-dopa (acetonide) -OH (1eq) was dissolved in dichloromethane at 0 ℃. Subsequently, 1-ethyl- (3-dimethylaminopropyl) carbonyldiimine hydrochloride (EDCI, 1-1.5 eq, preferably 1.2eq), 1-hydroxybenzotriazole (HOBt, 1-1.5 eq, preferably 1.2eq) and N, N-diisopropylethylamine (DIEA, 1-1.5 eq, preferably 1.2eq) are added in this order. Then in N₂A solution of N- (tert-butoxycarbonyl) -1, 4-diaminobutane (1.2eq) in methylene chloride was poured into the reaction mixture under ambient conditions, and the mixture was allowed to react at room temperature overnight. Adding appropriate amount of water to the reaction mixture, extracting the crude product into dichloromethane, washing the organic phase with saturated brine, anhydrous MgSO₄Dried and concentrated on a rotary evaporator. The crude product is purified by column chromatography to obtain a compound 1 which is a white solid with a yield of 68-75%.

Compound 1(1eq) was dissolved in tetrahydrofuran, and an aqueous solution of KOH (8eq) was injected into the reaction mixture under nitrogen protection, followed by reaction at room temperature for 5 hours. The solvent THF was removed by rotary evaporation, the crude product was extracted into ethyl acetate, the organic phase was washed with brine, dried over anhydrous MgSO4, and concentrated on rotary evaporator. Precipitation with petroleum ether gave the product as a white solid. The product (1eq) and Nalpha-fluorenylmethyloxycarbonyl-Nepsilon-tert-butyloxycarbonyl-L-lysine (Fmoc-Lys (Boc) -OH, 1-1.5 eq, preferably 1.2eq) were dissolved in dichloromethane at 0 ℃. Subsequently, EDCI (1-1.5 eq, preferably 1.2eq), HOBt (1-1.5 eq, preferably 1.2eq) and DIEA (1-1.5 eq, preferably 1.2eq) were added in this order and reacted at room temperature overnight. The reaction mixture was added with the appropriate amount of water, the crude product was extracted into dichloromethane, the organic phase was washed with saturated brine, dried over anhydrous MgSO4 and concentrated on rotary evaporator. The crude product is purified by column chromatography to obtain a compound 2 as a white solid with a yield of 70-83%.

Compound 2(1eq) was dissolved in tetrahydrofuran, and an aqueous solution of KOH (8eq) was injected into the reaction mixture under nitrogen protection, followed by reaction at room temperature for 5 hours. The solvent THF was removed by rotary evaporation, the crude product was extracted into ethyl acetate, the organic phase was washed with brine, dried over anhydrous MgSO4, and concentrated on rotary evaporator. Precipitation with petroleum ether gave the product as a white solid. The above product (1eq) and Fmoc-Dopa (acetonide) -OH (1-1.5 eq, preferably 1.05eq) were dissolved in dichloromethane at 0 ℃. Subsequently, EDCI (1-1.5 eq, preferably 1.2eq), HOBt (1-1.5 eq, preferably 1.2eq) and DIEA (1-1.5 eq, preferably 1.2eq) were added in this order and reacted at room temperature overnight. Adding appropriate amount of water to the reaction mixture, extracting the crude product into dichloromethane, washing the organic phase with saturated brine, anhydrous MgSO₄Dried and concentrated on a rotary evaporator. The crude product was purified by column chromatography to give compound 3 as a white solid in 66-76% yield.

The compound 3(1eq) was dissolved in tetrahydrofuran, and an aqueous solution of KOH (8eq) was poured into the reaction mixture under nitrogen protection, followed by reaction at room temperature for 5 hours. The solvent THF was removed by rotary evaporation, the crude product was extracted into ethyl acetate, the organic phase was washed with brine, dried over anhydrous MgSO4, concentrated by rotary evaporator and recrystallized from petroleum ether and dichloromethane to give compound DbaYKYThe intermediate is a white powdery solid, and the yield is 85-90%.

Containing DbaYKYSynthesis of end-group polymers

Further reaction of such polymers with Compound 4 or other initiator derived therefrom (claim 2) and removal of Boc and ketal protection gives the desired product containing DbaYKYA polymer of the group. Taking polylysine as an example, the synthetic route is shown below.

The polymerization was carried out in a nitrogen-protected glove box at room temperature. Neutralization of Boc-L-Lys-NCACompound 4 was added in a molar ratio of 1: 100, dissolving the mixture by using super-dry tetrahydrofuran, and reacting. After 48h of reaction at room temperature, the progress of the reaction was checked by TLC plates, and after the completion of the reaction, the reaction flask was taken out from the glove box and subjected to post-treatment of the polymerization reaction. Precipitation with tetrahydrofuran and petroleum ether gave a white solid, which was dried under vacuum. To remove the protecting group, the polymer was dissolved in trifluoroacetic acid (TFA): triisopropylsilane (TIS): h₂O is 95%: 2.5%: shaking the mixture in 2.5% deprotection solution at room temperature for 2 h. Excess trifluoroacetic acid was removed under a stream of nitrogen and a white fluffy solid was precipitated with methanol and ether and dried under vacuum to afford the deprotected polymer.

Versatile method for hydrogel functionalization

The formulation contains DbaYKYThe solvent of the terminal compound solution is various buffers such as Tris (hydroxymethyl) aminomethane (Tris-HCl), 4-hydroxyethylpiperazineethanesulfonic acid (HEPES), and Phosphate Buffer (PB), or an organic mixed solution, for example, Tris-HCl/MeOH ═ 4:6, preferably Tris-HCl buffer. The solvent has a pH of 5 to 9, preferably a pH of 8.5, and an ionic strength of 10mM to 500mM, preferably 100 mM. Immersing the hydrogel to be modified (swollen state or lyophilized state) into the prepared solution containing DbaYKYAnd (3) the solution of the terminal compound is kept for 6-48 hours, preferably 24 hours. And finally, sucking away the compound solution, replacing the compound solution with ultrapure water, replacing the ultrapure water once every 12 hours, and continuously replacing the ultrapure water for 5 times to obtain the functionalized hydrogel.

The functional hydrogels obtained by the above method can be widely used (including but not limited to) in the antibacterial and tissue engineering fields.

Example 1

DbaYKYSynthesis of intermediates

Synthesis of Compound 1

The compound Fmoc-DOPA (acetonide) -OH (10g, 21.8mmol) was placed in a 250mL round-bottomed flask at 0 deg.C and 80mL CH was added₂Cl₂Dissolved with stirring and cooled in an ice bath. EDCI (5.0g, 26.1mmol), 1-hydroxybenzotriazole (HOBt, 3.5g, 26.1mmol) and N, N-diisopropylethylamine (DIEA, 4.5mL, 26.1mmol) were added sequentially. N- (tert-Butoxycarbonyl) -1, 4-butanediamine (4.9g, 26.1mmol) was dissolved in 20mL CH₂Cl₂In N₂The mixture was added to the reaction flask with a syringe under protection. The reaction mixture was allowed to come to room temperature and stirred overnight. Extracting the crude product to CH₂Cl₂(300 mL. times.3), after 3 extractions, the organic phases were collected, washed together with saturated brine (100mL) and collected in an Erlenmeyer flask over anhydrous MgSO₄The product is dried and then filtered, and concentrated solution is obtained by rotary evaporation through a rotary evaporator. Chromatography on silica gel using PE and EtOAc as mobile phase gives 10.2g of pure product. The yield of this run was calculated to be 75%.

Characterization data for compound 1:¹H NMR(400MHz,CDCl₃)δ7.73(d,J＝7.6Hz,2H),7.52(t,J＝7.2Hz,2H),7.37(t,J＝7.6Hz,2H),7.28(d,J＝7.2Hz,2H),6.60(t,J＝7.6Hz,3H),6.42(br,1H),5.79(br,1H),4.75(br,1H),4.40(m,J＝10.4,7.2Hz,2H),4.25(br,1H),4.16(t,J＝6.8Hz,1H),3.34–2.80(m,6H),1.60(d,J＝4.4Hz,6H),1.42(s,9H),1.37(m,4H).¹³C NMR(100MHz,CDCl₃)δ171.14,156.15,147.60,146.37,143.73,141.24,129.60,127.72,127.11,125.13,125.05,121.85,119.98,117.88,109.45,108.11,79.14,67.05,56.48,47.06,40.01,39.12,38.56,28.44,27.38,26.38,25.80,25.79.HRESI-MS m/z calcd for C₃₆H₄₃N₃NaO₇[M+Na]⁺652.2999,found 652.2997.

synthesis of Compound 2

Compound 1(8.5g, 13.5mmol) was placed in a 250mL round bottom flask and dissolved by adding 60mL of THF with stirring. KOH (6.1g, 108.2mmol) was dissolved in 60mL H₂In O, in N₂The mixture was added to the reaction flask with a syringe under protection and stirred at room temperature overnight. The solvent THF was removed by rotary evaporation, the crude product was extracted into EtOAc (300 mL. times.3), and after 3 extractions the organic phases were collected together, washed with saturated brine (100mL) and collected in a conical flask over anhydrous MgSO₄The product was dried, filtered and concentrated to a minimum by rotary evaporation. 500mL of PE was added to a 1000mL flask containing the crude product, sonicated for 30 minutes, and filtered by suction to give a white solid which was used directly in the next step. The above solid (5.2g, 12.6mmol) and Fmoc-Lys (Boc) -OH (7.7g, 16.4mmol) were placed in a 250mL round bottom flask and 120mL CH was added₂Cl₂Dissolved with stirring and cooled in an ice bath. EDCI (3.2g, 16.4mmol), HOBt (2.2g, 16.4mmol) and DIEA (2.8mL, 16.4mmol) were added sequentially. The reaction mixture was allowed to come to room temperature and stirred overnight. Extracting the crude product to CH₂Cl₂(200 mL. times.3), after 3 extractions, the organic phases were collected, washed with saturated brine (100mL), collected in an Erlenmeyer flask, and then anhydrous MgSO₄And drying the product, filtering, and performing rotary evaporation by using a rotary evaporator to obtain a concentrated solution. Chromatography on silica gel using PE and EtOAc as mobile phase afforded 9.7g of pure product. The total yield of the above two experiments was calculated to be 83%.

Characterization data for compound 2:¹H NMR(400MHz,CDCl₃)δ7.74(d,J＝7.6Hz,2H),7.58(d,J＝6.8Hz,2H),7.38(t,J＝7.2Hz,2H),7.29(dd,J＝6.8,3.6Hz,2H),6.94(br,1H),6.56(d,J＝14.0Hz,4H),6.02(br,1H),4.85(br,2H),4.56(d,J＝6.4Hz,1H),4.44–4.24(m,2H),4.17(t,J＝6.8Hz,2H),3.34–2.78(m,8H),1.84–1.62(m,2H),1.56(d,J＝8.8Hz,6H),1.41(d,J＝6.4Hz,24H),1.26(d,J＝11.2Hz,2H).¹³C NMR(100MHz,CDCl₃)δ171.89,170.72,156.82,156.56,156.18,147.67,146.41,143.81,143.76,141.34,129.69,127.85,127.19,125.16,121.73,120.07,117.93,109.45,108.11,79.26,79.10,67.21,55.42,54.76,47.15,40.14,39.55,39.18,37.91,31.50,29.76,28.52,28.51,27.24,26.44,25.81,25.79,22.08.HRESI-MS m/z calcd for C₄₇H₆₃N₅NaO₁₀[M+Na]⁺880.4473,found 880.4474.

synthesis of Compound 3

Compound 2(9.5g, 11.1mmol) was placed in a 500mL round-bottom flask and dissolved with stirring by adding 120mL THF. KOH (5.0g, 88.8mmol) was dissolved in 120mL H₂In O, in N₂Adding into a reaction flask by a syringe under protection, addingThe mixture was stirred at room temperature overnight. The solvent THF was removed by rotary evaporation, the crude product was extracted into EtOAc (300 mL. times.3), and after 3 extractions the organic phases were collected together, washed with saturated brine (100mL) and collected in a conical flask, over anhydrous MgSO₄The product was dried, filtered and concentrated to a minimum by rotary evaporation. 500mL of PE was added to a 1000mL flask containing the crude product, sonicated for 30 minutes, and filtered by suction to give a white solid which was used directly in the next step. Dissolve the above solid (6.6g, 10.3mmol) and compound 2-3(5.0g, 10.8mmol) in 120mL CH₂Cl₂And cooled in an ice bath. EDCI (3.0g, 15.5mmol), HOBt (2.1g, 15.5mmol) and DIEA (2.7mL, 15.5mmol) were then added. The reaction mixture was allowed to come to room temperature and stirred overnight. Extracting the crude product to CH₂Cl₂(200 mL. times.3), after 3 extractions, the organic phases were collected, washed with saturated brine (100mL), collected in an Erlenmeyer flask, and then anhydrous MgSO₄The product is dried and then filtered, and concentrated solution is obtained by rotary evaporation through a rotary evaporator. The concentrate was placed in a 1000mL round bottom flask and 500mL EtOAc was added to dissolve the precipitate, sonicated for 30 minutes and filtered by suction to give 9.0g of a white solid. The total yield of the above two experiments was calculated to be 76%.

Characterization data for compound 3:¹H NMR(400MHz,DMSO-d6)δ8.05(d,J＝7.6Hz,1H),7.95–7.81(m,4H),7.61(dd,J＝13.2,6.8Hz,3H),7.40(td,J＝7.6,3.6Hz,2H),7.33–7.23(m,,2H),6.82(d,J＝1.2Hz,1H),6.78–6.62(m,6H),6.59(dd,J＝8.0,1.2Hz,1H),4.43–4.01(m,6H),3.10–2.55(m,10H),1.52–1.41(m,12H),1.52–1.41(m,2H),1.41–1.25(m,24H),1.25–1.14(m,2H).¹³C NMR(100MHz,DMSO-d6)δ171.60,171.17,170.43,155.85,155.59,155.53,146.60,145.37,145.29,143.76,143.72,140.65,131.31,130.64,127.61,127.08,125.37,125.27,121.79,121.67,120.11,117.45,109.47,109.32,107.63,77.35,77.32,65.69,56.29,54.13,52.69,46.57,38.23,37.71,37.08,31.84,29.24,28.28,26.79,26.30,25.50,22.60.HRESI-MS m/z calcd for C₅₉H₇₆N₆NaO₁₃[M+Na]⁺1099.5368,found1099.5359.

DabYKYintermediate ofSynthesis of the body

Compound 3(1g, 0.9mmol) was placed in a 50mL round-bottom flask, 10mL THF was added and dissolved with stirring, KOH (0.4g, 7.4mmol) was dissolved in 10mL H₂In O, in N₂The mixture was added to the reaction flask with a syringe under protection and stirred at room temperature overnight. By using CH₂Cl₂Extracting, adding PE for precipitation, and using CH₂Cl₂Recrystallization from PE 1:2 gave 714mg of pure product. The yield of this run was calculated to be 90%.

Characterization data for compound DabYKY intermediate:¹H NMR(400MHz,CDCl₃)δ7.84(s,1H),6.88(s,1H),6.75–6.39(m,7H),4.84(d,J＝36.4Hz,2H),4.51(dd,J＝14,6.8Hz,1H),4.28(d,J＝5.6Hz,1H),3.51(s,1H),3.33–2.80(m,9H),2.70-2.50(m,1H),2.40–1.95(m,3H),1.78(dd,J＝13.2,7.2Hz,1H),1.61(d,J＝18.8Hz,12H),1.41(s,24H).¹³C NMR(100MHz,CDCl₃)δ175.30,171.49,170.75,156.22,147.89,147.54,146.50,146.37,130.33,129.76,128.72,121.92,121.87,120.03,118.04,117.99,114.93,109.49,109.23,108.22,108.15,79.11,56.25,54.58,53.32,40.43,40.19,39.20,37.65,31.48,29.76,29.60,28.52,27.23,26.45,25.92,25.83,25.80,22.74.HRESI-MS m/z calcd for C₄₄H₆₆N₆NaO₁₁[M+H]⁺854.4790,found855.4869.

example 2

Synthesis of ATRP polymerization initiator

Dba with protective groupYKYThe terminal intermediate (320mg, 0.37mmol) and 2-bromo-2-methylpropionic acid (75mg, 0.45mmol) were placed in a 50mL round bottom flask, and 8mL CH was added₂Cl₂Stirring to dissolve. Then at N₂EDCI (108mg, 0.56mmol), HOBt (76mg, 0.56mmol) and DIEA (97. mu.L, 0.56mmol) were added successively with protection. The mixture was stirred at room temperature overnight. Extracting the crude product to CH₂Cl₂(50 mL. times.3), after 3 extractions, the organic phase was separatedCollected together, washed with saturated brine (50mL) and collected in an erlenmeyer flask over anhydrous MgSO₄The product is dried and then filtered, and concentrated solution is obtained by rotary evaporation through a rotary evaporator. Subjecting to silica gel column chromatography with CH₂Cl₂And MeOH as the mobile phase to give 310mg of pure product. The yield of this experiment was calculated to be 83%. Compound characterization data:¹H NMR(600MHz,CD₃OD)δ6.74–6.54(m,6H),4.67(dd,J＝6.0,3.2Hz,1H),4.50(dd,J＝9.6,5.2Hz,1H),4.43–4.34(m,1H),3.27–2.83(m,10H),1.86(d,J＝12.8Hz,6H),1.80–1.72(m,2H),1.71–1.62(m,2H),1.62–1.50(m,12H),1.50–1.28(m,24H).HRESI-MS m/z calcd for C₄₈H₇₀N₆O₁₂Br[M-H]^-1001.4235,found 1001.4238.

example 3

Synthesis of RAFT polymeric chain transfer reagent

Dba with protective groupYKYThe end group intermediate (320mg, 0.37mmol) and S-thiobenzoylthioglycolic acid (96mg, 0.45mmol) were placed in a 50mL round bottom flask and 8mL CH was added₂Cl₂Stirring to dissolve. Then at N₂EDCI (108mg, 0.56mmol), HOBt (76mg, 0.56mmol) and DIEA (97. mu.L, 0.56mmol) were added successively under protection. The mixture was stirred at room temperature overnight. Extracting the crude product to CH₂Cl₂(50 mL. times.3), after 3 extractions, the organic phases were collected, washed together with saturated brine (50mL) and collected in an Erlenmeyer flask, and then dried over anhydrous MgSO₄And drying the product, filtering, and performing rotary evaporation by using a rotary evaporator to obtain a concentrated solution. Subjecting to silica gel column chromatography with CH₂Cl₂And MeOH as the mobile phase to give 322mg of pure product. The yield of this experiment was calculated to be 82%. Compound characterization data:¹H NMR(400MHz,CDCl₃)δ7.98(d,J＝7.6Hz,2H),7.69–7.28(m,5H),7.10–6.75(m,2H),6.72–6.37(m,6H),4.79–4.02(m,7H),3.33–2.73(m,10H),1.81–1.54(m,14H),1.43(s,24H),1.17(s,2H).HRESI-MS m/z calcd for C₅₃H₇₁N₆O₁₂S₂[M-H]^-1047.4571,found 1047.4574.

example 4

Synthesis of ROP initiator

The starting material CP (4.0g, 36mmol) was placed in a 500mL round-bottom flask, anhydrous tetrahydrofuran (THF, 200mL) was added and dissolved with stirring, and the solution was cooled to-78 ℃. Lithium bis- (trimethylsilyl) amide (LiHMDS, 7.8g, 46.8mmol) was dissolved in 47mL THF, this solution was added to the reaction flask under nitrogen, the mixed solution was stirred for 10 minutes, succinic anhydride (10g, 108mmol) was dissolved in THF (20mL) solution and injected dropwise with syringe. After 1 hour, the reaction was warmed to-30 ℃ and 1M NaHSO₄Quenching and adjusting the pH to 2-3. After removing most of the THF by rotary evaporation, the crude product was extracted into ethyl acetate (EtOAc, 200 mL. times.3), and after 3 extractions were completed, the organic phases were collected together, washed with saturated brine (100mL) and collected in a conical flask, over anhydrous MgSO₄The product was dried and filtered and rotary evaporated by a rotary evaporator to give a concentrated solution for further use.

The solid was placed in a 500mL round bottom flask and anhydrous dichloromethane (CH) was added₂Cl₂300mL) was dissolved with stirring, and then 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDCI, 10.4g, 54mmol) and N-hydroxysuccinimide (NHS, 6.2g, 54mmol) were added in that order. The mixture was stirred at room temperature overnight and the progress of the reaction was monitored by Thin Layer Chromatography (TLC) spot plate. Extracting the crude product to CH₂Cl₂(200 mL. times.3), after 3 extractions, the organic phases were collected, washed with saturated brine (100mL) and collected in an Erlenmeyer flask, and then dried over anhydrous MgSO₄After drying the productFiltering, and rotary evaporating by a rotary evaporator to obtain a concentrated solution. Chromatography on silica gel using Petroleum Ether (PE) and EtOAc as mobile phase afforded 7.9g of NHS-CP β pure product in 71% yield. Compound characterization data:¹H NMR(400MHz,CDCl₃)δ4.42(t,J＝4.8Hz,1H),3.53(dd,J＝8.0,4.4Hz,1H),3.15–3.03(m,2H),3.01–2.92(m,2H),2.81(s,4H),2.26(dd,J＝13.6,5.2Hz,1H),2.09(dd,J＝12.0,4.8Hz 1H),1.93–1.83(m,1H),1.67–1.50(m,2H),1.49–1.36(m,1H).¹³C NMR(100MHz,CDCl₃)δ169.03,168.20,167.89,56.88,54.38,31.13,28.58,25.90,25.67,25.22,22.82.HRESI-MS m/z calcd for C₁₄H₁₆N₂NaO₆[M+Na]⁺331.0906,found 331.0907.

dba with protective groupYKYThe end-group intermediate (500mg, 0.585mmol) was dissolved in 10mL of anhydrous CH₂Cl₂Then in N₂NHS-CP β (198.3mg, 0.643mmol) and triethylamine (TEA, 8.1 μ L, 0.0585mmol) were added with protection. The mixture was stirred at room temperature for 3 hours. Extracting the crude product to CH₂Cl₂(50 mL. times.3), after 3 extractions, the organic phases were collected, washed together with saturated brine (50mL) and collected in an Erlenmeyer flask over anhydrous MgSO₄The product is dried and then filtered, and concentrated solution is obtained by rotary evaporation through a rotary evaporator. Column chromatography on silica gel with cyclohexane and EtOAc as the mobile phase afforded 540mg of pure product. The yield of this experiment was calculated to be 88%. Compound characterization data:¹H NMR(400MHz,CDCl₃)δ8.04–7.30(m,2H),7.12–6.73(m,2H),6.57(d,J＝29.6Hz,6H),5.23–4.86(m,2H),4.66(br,2H),4.37(m,1H),4.18(m,1H),3.52(dd,J＝6.8,4.4Hz,1H),3.33–2.68(m,12H),2.62–2.36(m,2H),2.23–1.95(m,2H),1.94–1.69(m,2H),1.66–1.49(m,16H),1.39(s,24H).¹³C NMR(100MHz,CDCl₃)δ171.90,171.13,171.05,170.51,168.12,168.06,156.23,148.00,147.87,147.51,146.74,146.63,146.18,145.99,130.82,130.39,129.37,121.73,121.25,118.15,117.72,109.41,109.15,108.35,108.28,107.78,78.91,56.86,56.70,54.40,40.35,39.26,37.43,32.00,29.77,28.73,28.54,26.94,26.53,25.90,25.81,22.87,22.77,14.21.HRESI-MS m/z calcd for C₅₄H₇₇N₇NaO₁₄[M+Na]⁺1070.5426,found1070.5416.

example 5

Containing DbaYKYSynthesis of terminal quaternized Poly (N, N-dimethylaminoethyl methacrylate) (QPDMAEMA).

The polymer is obtained by Atom Transfer Radical Polymerization (ATRP) using dimethylaminoethyl methacrylate (DMAEMA) as a monomer, followed by quaternization. The polymerization was carried out in a nitrogen-protected glove box at room temperature. The ultra-dry solvent THF was bubbled under a stream of nitrogen for 15 minutes to remove oxygen from the solvent before the polymerization reaction was carried out.

The polymerization process was carried out as follows: DMAEMA (340. mu.L, 2mmol), ATRP initiator (10mg, 0.01mmol), and 1,1,4,7,10, 10-hexamethyltriethylenetetramine (HMTETA, 6.5. mu.L, 0.024mmol) were placed in a reaction flask equipped with a magnetic stirrer and dissolved with stirring by adding extra dry THF (200. mu.L). CuCl (2.7mg, 0.02mmol) and CuCl were added sequentially₂(0.4mg, 0.004 mmol). Samples were taken at various time periods during the polymerization process by GPC to analyze polymer molecular weight Mn and molecular weight distribution. After 24h, the polymerization was terminated by exposing the reaction to air. The mixture was then diluted with 500 μ L THF and passed through a neutral alumina column to remove the catalyst. The above oily substance was dissolved in 1mL of THF, and then 45mL of cold Petroleum Ether (PE) was slowly added to the mixture to precipitate a white powdery polymer. The centrifuge tube was centrifuged at 4500rpm for 3min and the supernatant was removed, and then the polymer was collected and dried under a stream of nitrogen. After repeating the dissolution/precipitation process three times, the white solid was dried under vacuum.

50mg of the above polymer was dissolved in 1mL of MeCN, then excess 1-bromooctane (550. mu.L) was added and the reaction mixture was stirred at 55 ℃ for 24 h. After removal of the solvent by rotary evaporation, the oil obtained is dissolved in0.5mL MeOH, then 45mL cold diethyl ether (Et)₂O) was slowly added to precipitate the polymer, this operation was repeated three times, and vacuum dried to obtain the quaternized polymer. To remove the protecting group, the volume ratio was trifluoroacetic acid (TFA): triisopropylsilane (TIS): h₂O95%: 2.5%: the 2.5% solution was added to a 50mL centrifuge tube containing the polymer, which was gently shaken for 2h to dissolve the polymer completely. Excess trifluoroacetic acid is removed under a stream of nitrogen and the mixture is dissolved in 0.5mL MeOH, followed by slow addition of cold 45mL Et₂O to precipitate a white fluffy solid. The precipitate was collected by centrifugation and washed with water and brine₂And (4) drying under flowing. After repeating the dissolution-precipitation cycle three times, drying under vacuum gave the final polymer.

Example 6

Containing DbaYKYSynthesis of end-group beta DM CP polymers

The polymerization was carried out at room temperature in a nitrogen-protected glove box. DM monomer (27.4mg, 0.12mmol) and CP monomer (20.0mg, 0.18mmol) were first added to the reaction flask and dissolved with 1mL of extra dry THF with stirring. ROP initiator (3.1mg, 0.003mmol) was dissolved in 0.5mL THF, LiHMDS (1.3mg, 0.0075mmol) was also dissolved in 0.5mL THF and added to the reaction flask in sequence, the mixture was stirred for 4h, the progress of the reaction was checked by TLC plate, after completion of the reaction, the reaction flask was removed from the glove box, the liquid in the reaction flask was transferred to a 50mL centrifuge tube, 1mL THF was added for dissolution, and then 45mL cold PE was slowly added to the mixture to precipitate a powdery polymer. The centrifuge tube was centrifuged at 4500rpm for 3min and the supernatant was removed before the polymer was collected and dried under a stream of nitrogen. After repeating the dissolution/precipitation process five more times, the solid was dried under vacuum.

To remove the protecting group, the volume ratio was TFA: and (3) TIS: h₂O is 95%: 2.5%: 2.5% of the deprotecting solution was added to 50mL of the solution containing the above polymerThe polymer was completely dissolved in the heart tube, and the tube was gently shaken for 2 h. Excess TFA was removed under a stream of nitrogen, the mixture was dissolved in 0.5mL MeOH, and then cold 45mL Et was added slowly₂O to precipitate a white fluffy solid. The precipitate was collected by centrifugation and washed with water and brine₂And (4) drying under flowing. After repeating the dissolution-precipitation cycle three times, drying under vacuum gave the deprotected polymer.

Example 7

Containing DbaYKYTerminal QPDMAEMA modified PEG hydrogel

Preparation of polyethylene glycol (PEG) hydrogel: polyethylene glycol diacrylate (PEGDA, molecular weight 2000) was dissolved in ultrapure water at a concentration of 20 wt%, the photoinitiator 2-hydroxy-4' - (2-hydroxyethoxy) -2-methylpropiophenone (I2959) was dissolved in ethanol at a concentration of 10 wt% to prepare an initial solution, and the initial solution of the photoinitiator was mixed with the PEGDA solution so that the final concentration of the photoinitiator was 0.1 wt%. Placing the mixed solution in a mold, and applying ultraviolet rays (365nm,50 mw/cm)²) After 5 minutes, a hydrogel was formed.

MeOH and 100mM Tris-HCl buffer were mixed at a volume ratio of 4:6, and the polymer QPDMAEMA was dissolved in the mixed solution at a concentration of 4 mg/mL. Wiping the surface of the PEG hydrogel with dust-free paper to remove water, dripping the polymer solution on the surface of the hydrogel to ensure that the whole surface is covered with the liquid drop, flushing the surface with ultrapure water after 24h, immersing the modified hydrogel in the ultrapure water for 12h, changing the water every 3h to remove the polymer which is not modified on the surface of the hydrogel, and performing N₂And (5) drying by blowing down to obtain the QPDMAEMA modified PEG hydrogel surface.

The successful modification of the hydrogel by the polymer was characterized by infrared spectroscopy. The infrared spectrum of the QPDMAEMA modified PEG hydrogel compared with the unmodified PEG hydrogel is 3350cm^-1Shows a stretching vibration peak of OH and is 2925cm^-1And 2855cm^-1There are C-H stretching vibration peaks, which are the groups on the polymer chain. As a control, Dba was absentYKYQuaternized poly (N, N-dimethylaminoethyl methacrylate) modified with an adhesive groupPEG hydrogel does not appear to be 3350cm^-1、2925cm^-1And 2855cm^-Indicating that the polymer was not able to modify the hydrogel and was not bound to the hydrogel.

Example 8

Containing DbaYKYSterilization testing of terminal QPDMAEMA-modified PEG hydrogels

The bacteria used in this experiment were the gram-positive bacteria Staphylococcus aureus (s. aureus, ATCC 6538) and the gram-negative bacteria Escherichia coli (e. coli, ATCC 25922). The bacteria were cultured in an Erlenmeyer flask containing LB liquid medium, and the Erlenmeyer flask was placed on a shaker (150rpm) at 37 ℃ for 10 hours. After the culture is finished, pouring the bacterial liquid in the conical flask into a 50mL centrifugal tube, centrifuging at 4000rpm for 5min, collecting the precipitate, adding 5mL PBS, shaking again, dispersing uniformly, centrifuging again, repeatedly using the PBS to disperse the bacterial liquid for three times, collecting the bacterial liquid, and using an enzyme-linked immunosorbent assay (ELISA) reader to read OD values to quantify the colony number. The bacterial solution was diluted to 10 with PBS⁶CFU/mL is ready for use. QPDMAEMA-modified PEG hydrogel surfaces were placed in 24-well plates with unmodified PEG hydrogel surfaces as controls. 80 μ L of the above bacteria is concentrated to 10%⁶CFU/mL of the bacterial solution was applied to the surface of each hydrogel, PBS was added to a blank well plate for humidity control, and the 24-well plate was placed in a 37 ℃ mold incubator for culture. After 2.5h incubation, the well plate was removed, 1920. mu.L PBS was added to dilute the plate, the plate was sonicated for 3min, mixed for 2min under the mixer, then 30. mu.L was removed with a pipette and added to the previously prepared LB agar medium to coat it evenly, and incubated overnight in a 37 ℃ mold incubator. The colonies were counted by plate counting method and the experimental group was designated C_polymerBlank control is denoted C_control. The antibacterial activity (bacterial killing rate) of the surface of different substrates is calculated by the following formula:

when the polymer QPDMAEMA is dripped on the surface of the PEG hydrogel, the surface of the hydrogel modified by the polymer is against gram-positive bacteriaThe S.aureus and the gram-negative bacteria E.coli have high-efficiency sterilization effects, and the sterilization rate is more than 99 percent. Number of colonies on surface of unmodified PEG hydrogel and C_controlRather, there is no bactericidal capacity.

Example 9

Containing DbaYKYPHEMA hydrogel modified by terminal group beta DM-CP polymer

Preparation of poly (2-hydroxyethyl methacrylate) (PHEMA) hydrogel: hydroxyethyl methacrylate monomer (HEMA, 50 wt%), crosslinker N, N' -methylenebisacrylamide (MBAA, 0.8 wt%) was added to the mixed solvent (ethylene glycol/ethanol/water ═ 1.5:1: 1.5). The photoinitiator I2959 was dissolved in ethanol at a concentration of 10 wt% to prepare an initial solution, and the initial solution of the photoinitiator was added to the above mixed solution to give a final concentration of 0.1 wt% of the photoinitiator. The mixed solution was placed in a mold, and gelled for 12 minutes by ultraviolet rays (365nm,50mw/cm2) to form a hydrogel.

Will contain DbaYKYEnd-group beta DM CP Polymer was dissolved at a concentration of 4mg/mL in 100mM Tris-HCl buffer. Wiping the surface of the PHEMA hydrogel with dust-free paper to remove water, dropwise adding the polymer solution onto the surface of the hydrogel to enable the liquid drops to be paved on the whole surface, washing the surface with anhydrous ethanol and ultrapure water in sequence after 24 hours, immersing the modified hydrogel in the ultrapure water for 12 hours, changing water every 3 hours to remove the polymer which is not modified on the surface of the hydrogel, and removing the polymer on the surface of the hydrogel by N₂And blowing down and drying to obtain the beta DM-CP polymer modified hydrogel surface.

The successful modification of the hydrogel by the polymer was characterized by infrared spectroscopy. Compared with the unmodified PHEMA hydrogel, the beta DM CP polymer modified PHEMA hydrogel has an infrared spectrum shown at 1650cm^-1The stretching vibration peak of NH appears, which is the peak of the radicals on the polymer chain. As a control, there was no DbaYKYAttachment group beta DM CP Polymer modified PEG hydrogel with no appearance of 1650cm^-1Indicating that the polymer failed to modify the hydrogel and did not bind to the hydrogel.

Example 10

Containing DbaYKYBeta D of terminal groupsCell adhesion test on PHEMA hydrogel surface modified by CP (carboxy-terminated polyethylene) polymer

Incubate at 37 ℃ in DMEM medium supplemented with 10% Fetal Bovine Serum (FBS), 100. mu.g/mL streptomycin, 100U/mL penicillin, and 2mmol L-glutamine (5% CO)₂95% humidity) were cultured in NIH 3T3 fibroblasts. When the coverage of the cells in the culture dish reached about 90%, the cells were diluted to 3X 10 with DMEM medium⁵cell/mL is ready for use. Will contain DbaYKYEnd group beta DM CP Polymer-modified PHEMA hydrogel surface was placed in a 24-well plate with the unmodified PHEMA hydrogel surface as a control. 50 μ L of the suspension was added at a concentration of 3X 10⁵cell/mL cell suspension was dropped onto the hydrogel surface and the 24-well plate was placed in a 37 ℃ incubator for culture. And (3) taking out the pore plate after 2h, adding 1mL of DMEM medium into each pore, continuously placing the pore plate in the incubator for culturing, taking out the pore plate after 24h, sucking out the DMEM medium in the pore plate by using a pipette gun, and washing the pore plate twice by using PBS. The adhesion and growth of the cells on the surface were observed using Live-Dead Cell Staining Kit (Live-Dead Cell Staining Kit). And dropwise adding the prepared 30 mu L of live-dead cell staining reagent on the surface of each hydrogel for incubation for 10 min. After the incubation, the surface was washed twice with PBS and the staining of the cells was directly observed under an inverted fluorescence microscope.

Since PHEMA hydrogel itself has an antifouling effect, there is little fibroblast adhesion on the unmodified hydrogel. Conversely, fibroblasts can adhere to and spread in a medium containing DbaYKYCP polymer modified PHEMA hydrogels, in sharp contrast to unmodified hydrogels.

Example 11

Containing DbaYKYEnd-group beta DM CP Polymer modified HEMA hydrogel internal cell adhesion test

To examine the effect of modifying the polymer inside the hydrogel, the lyophilized PHEMA hydrogel was immersed in a solution containing Dba at a concentration of 4mg/mLYKYCP Polymer solution, after 24h, the modified hydrogel was immersed in ultrapure water for 12h, and the water was changed every 2h to remove the unmodified hydrogel surfaceIn N of₂Flow-down blow-dry, then cut the modified PHEMA hydrogel from the middle with a knife to expose DbaYKYBeta DM of the end group the internal cross section of PHEMA hydrogel modified by CP polymer, and the cell adhesion experiment is carried out on the cross section.

When the PHEMA hydrogel modified with the β DM: CP polymer was cut into two parts from the middle and fibroblasts were seeded on the inner cross section of the PHEMA hydrogel, the results showed that both inner surfaces showed excellent cell adhesion, while the unmodified hydrogel showed little fibroblast adhesion, indicating that the gel was obtained by DbaYKYThe simple one-step modification of the terminal polymer effectively functionalizes the surface and the interior of the hydrogel.

The successful modification of the hydrogel by the polymer was characterized by infrared spectroscopy. The infrared spectrum of the beta DM CP polymer modified PHEMA hydrogel is shown at 1650cm^-1And a stretching vibration peak of NH appears, which is consistent with the infrared characterization of the modified PHEMA hydrogel surface.

Example 12

Containing DbaYKYTerminal group beta DM CP polymer modified PVA hydrogel

Preparation of PVA hydrogel: PVA was added to ultrapure water at a concentration of 14% by weight and dissolved by heating and stirring in an oil bath at 95 ℃ for 4 hours. And after the PVA is completely dissolved, placing the solution in an ultrasonic cleaning machine to remove bubbles for 30min to obtain a uniform and transparent solution. The PVA solution was placed in a mold and frozen at-18 ℃ for 16 hours, thawed at room temperature for 8 hours, and the hydrogel was formed after 3 freeze-thaw cycles.

Will contain DbaYKYCP Polymer was dissolved at a concentration of 4mg/mL in 100mM Tris-HCl buffer. Wiping water on the surface of PVA hydrogel by using a piece of dust-free paper, dripping the polymer solution on the surface of the hydrogel to ensure that the whole surface is paved by the liquid drops, washing the surface by using anhydrous ethanol and ultrapure water in sequence after 24h, immersing the modified hydrogel in the ultrapure water for 12h, changing the water every 3h to remove the polymer which is not modified on the hydrogel, and carrying out N-phase polymerization on the polymer solution₂Drying by flowing down to obtain DbaYKYTerminal beta DM: CPThe polymer modifies the hydrogel surface.

The successful modification of the hydrogel by the polymer was characterized by infrared spectroscopy. Compared with the unmodified PVA hydrogel, the infrared spectrum of the beta DM CP polymer modified PHEMA hydrogel is shown at 1650cm^-1The stretching vibration peak of NH appears, which is the peak of the radicals on the polymer chain.

Example 13

Containing DbaYKYTerminal group beta DM CP Polymer modified PVA-promoted wound healing function test to evaluate the role of the beta DM CP Polymer modified PVA hydrogel in wound healing in vivo, a mouse whole cortex wound model was established. Sterile Sprague-Dawley (SD) female rats were raised for 8-10 weeks and the body weight reached 200-250g for experiments in which the rats were randomly assigned to PVA hydrogels (control group) and contained DbaYKYTwo groups of PVA hydrogel modified by CP polymer. Experimental rats were anesthetized with 1% sodium pentobarbital (5mg/kg) before the procedure, the excess hair on the back of the rats was shaved off using a pet razor, the depilatory cream was applied to the backs of the rats, and the residual depilatory cream was removed after 10min of action. The back of the rat is cleaned and wiped by dipping a cotton swab in physiological saline, and finally the back of the rat is disinfected by alcohol. Full-thickness circular wounds of 8mm diameter were made with a biopsy punch, PVA hydrogel of 8mm diameter and beta DM: CP Polymer modified PVA hydrogel were placed directly on each wound, and the wound surface was then covered with a clear dressing (3M Tegaderm Film) and a medical bandage. On days 0, 3, 5, 7,10, the wounds were photographed and the area of the wounds was analyzed with Image-J. The calculation formula of the wound healing degree is as follows:

wherein A is₀And A_tWound area on day 0 and t, respectively. Mice were sacrificed on day 10, wound skin tissue was removed, fixed in 4% paraformaldehyde and left overnight, stained with hematoxylin-eosin (H)&E) Staining tissue sections were analyzed for staining.

PVA hydrogel and Dba-containing PVA hydrogelYKYCP polymer modified PVA hydrogel was placed directly on each full-thickness wound, and the healing process of the wound was carefully monitored. Almost healing of the wound was observed on day 10 for the polymer-modified PVA hydrogel group, while a larger area of scab remained in the PVA hydrogel group. Hematoxylin and eosin (H) were used on day 10&E) The microscopic effect of wound healing was observed by staining, the polymer-modified PVA hydrogel group developed an intact epidermis and observed hair follicles, and the neogenetic tissue was substantially identical to normal skin and was substantially completely repaired, but the epidermis of the PVA hydrogel group was incomplete and accompanied by scars, and a large number of inflammatory cells were present in the neogenetic tissue, which was clearly different from the surrounding normal tissue.

Example 14

Containing DbaYKYEnd group beta DM CP Polymer modification of various surfaces

Taking a gold surface as an example, containing DbaYKYDissolving the terminal group beta DM CP polymer in 100mM Tris-HCl buffer solution at the concentration of 4mg/mL, dropwise adding the solution on the gold-plated surface cleaned by ultraviolet ozone to ensure that the solution is dripped on the whole surface, washing the surface by ultrapure water, ethanol and ultrapure water in sequence after 24 hours, and washing the surface by N₂Drying by flowing down to obtain DbaYKYEnd group beta DM-gold surface modified by CP Polymer. The characterization by XPS shows that the unmodified gold surface has only obvious Au peak, and Dba shows thatYKYEnd group β DM: CP polymer modified surface showed distinct C, O and N peaks, demonstrating successful modification.

Example 15

Containing DbaYKYSterilization test of terminal QPDMAEMA-modified various surfaces

The bacteria used in this experiment were the gram-positive bacteria Staphylococcus aureus (s. aureus, ATCC 6538) and the gram-negative bacteria Escherichia coli (e. coli, ATCC 25922). The bacteria were cultured in an Erlenmeyer flask containing LB liquid medium, and the Erlenmeyer flask was placed on a shaker (150rpm) at 37 ℃ for 10 hours. After the culture is finished, pouring the bacterial liquid in the conical flask into a 50mL centrifugal tube, centrifuging at 4000rpm for 5min, collecting the precipitate, adding 5mL PBS, shaking again, dispersing uniformly, and centrifuging againAnd repeatedly centrifuging the PBS dispersed bacteria liquid for three times, collecting the bacteria liquid, and reading the OD value by using an enzyme-labeling instrument to quantify the colony number. The bacterial liquid is diluted to 10 by PBS⁶CFU/mL is ready for use. Place QPDMAEMA-modified various surfaces into 24-well plates, including Ti₂O, gold, glass, silicon, polyurethane and polyetheretherketone surfaces. The various types of surfaces without modification served as controls. 80 μ L of the above bacteria is concentrated to 10%⁶CFU/mL of the bacterial solution was applied to the surface of each substrate, PBS was added to a blank well plate for humidity control, and the 24-well plate was placed in a 37 ℃ mold incubator for culture. After 2.5h incubation, the well plate was removed, 1920. mu.L PBS was added to dilute the plate, the plate was sonicated for 3min, mixed for 2min under the mixer, then 30. mu.L was removed with a pipette and added to the previously prepared LB agar medium to coat it evenly, and incubated overnight in a 37 ℃ mold incubator. The colonies were counted by plate counting method and the experimental group was marked C_polymerBlank control is denoted C_control. The antibacterial activity (bacterial killing rate) of the surface of different substrates is calculated by the following formula:

when the polymer QPDMAEMA is dripped on various surfaces, all the surfaces modified by the polymer have high-efficiency bactericidal effects on gram-positive bacteria S.aureus and gram-negative bacteria E.coli, and the bactericidal rate is more than 99 percent. Number of unmodified surface colonies of each type and C_controlRather, there is no bactericidal capacity.

Example 16

Containing DbaYKYEnd group beta DM CP Polymer modified PTFE surface cell adhesion test

Incubate at 37 ℃ in DMEM medium supplemented with 10% Fetal Bovine Serum (FBS), 100. mu.g/mL streptomycin, 100U/mL penicillin, and 2mmol L-glutamine (5% CO)₂95% humidity) to culture NIH 3T3 fibroblasts. When the coverage of the cells in the culture dish reached about 90%, the cells were diluted to 3X 10 with DMEM medium⁵cell/mL is ready for use. Will contain DbaYKYTerminal groupCP polymer modified PTFE surface was placed in a 24-well plate with an unmodified PTFE surface as a control. 50 μ L of the suspension was added at a concentration of 3X 10⁵cell/mL of the cell suspension was dropped onto the PTFE surface, and the 24-well plate was placed in a 37 ℃ incubator for culture. And (3) taking out the pore plate after 2h, adding 1mL of DMEM medium into each pore, continuously placing the pore plate in the incubator for culturing, taking out the pore plate after 24h, sucking out the DMEM medium in the pore plate by using a pipette gun, and washing the pore plate twice by using PBS. The adhesion and growth of cells on the surface were observed using Live-Dead Cell Staining Kit (Live-Dead Cell Staining Kit). And dropwise adding the prepared 30 mu L of live-dead cell staining reagent on the surface of each hydrogel for incubation for 10 min. After the incubation, the surface was washed twice with PBS, and the staining of the cells was directly observed under an inverted fluorescence microscope.

Since the PTFE hydrogel itself has an anti-adhesion function, there is little fibroblast adhesion on the unmodified PTFE surface. Conversely, fibroblasts can adhere to and spread in a medium containing DbaYKYCP polymers modified PTFE surface, in sharp contrast to the unmodified surface.

All documents referred to herein are incorporated by reference into this application as if each were individually incorporated by reference. Furthermore, it should be understood that various changes and modifications of the present invention can be made by those skilled in the art after reading the above teachings of the present invention, and these equivalents also fall within the scope of the present invention as defined by the appended claims.

Claims

1. A compound or polymer containing phenolic adhesion groups, wherein the phenolic adhesion groups are comprised of repeating units represented by formulas I and II:

in the formula, R is a positive charge side group and is a side group containing primary amine, secondary amine or tertiary amine; each chiral center may be S-type or R-type.

2. A phenolic adhesion group-containing compound or polymer as claimed in claim 1 wherein the phenolic adhesion group is:

3. the phenolic adhesion group-containing compound or polymer of claim 1, wherein the polymer has the structure of formula V:

wherein R, x and y are as defined in claim 1;

-/- (-in the arrangement in random, block or alternate form;

R₂h, C1-C18 alkyl, amino-substituted C1-C18 alkyl, hydroxyl, amino, carboxyl, aldehyde, C2-C18 alkenyl, sulfydryl, azide, C2-C18 alkynyl, and o-dithiopyridyl (OPSS); or a group containing maleimide, biotin, adamantane, cyclodextrin; or a polymer chain selected from amino acid polymer chains and carbon chain polymer chains.

4. The phenolic adhesion group-containing polymer of claim 1, wherein the polymer is selected from the group consisting of:

in each formula, each n is independently an integer of 1 to 1000.

5. The method of any of claims 1-4, wherein the method of preparation initiates polymerization by an initiator selected from the group consisting of:

6. a functional hydrogel comprising a hydrogel matrix modified at the surface and/or interior with the phenolic adhesion group-containing polymer of claim 1.

7. The functional hydrogel of claim 6, wherein said hydrogel matrix is selected from the group consisting of: polyethylene glycol (PEG) hydrogel, polyvinyl alcohol (PVA) hydrogel, Polyhydroxyethylmethacrylate (PHEMA) hydrogel, Polysulfonobetaine (PSBMA) hydrogel, polycarboxybetaine (pcbmma) hydrogel, alginic Acid (ALG) hydrogel, Hyaluronic Acid (HA) hydrogel, dextran hydrogel, chitosan hydrogel.

8. The method of claim 6, wherein the method comprises the step of immersing the hydrogel in a solution of the polymer comprising phenolic adhesive groups to obtain a functional hydrogel.

9. Use of a functional hydrogel according to claim 6 for the preparation of an antibacterial material or for the preparation of a tissue engineering scaffold.

10. A method for general surface modification and functionalization of specific end group, characterized in that the specific end group has the following structure: