Sulfonyl zwitterionic polymer with antibacterial function and preparation method and application thereof

Abstract

Sulfonyl zwitterionic polymers with antibacterial properties are formed by polymerizing sulfonyl zwitterionic monomers with crosslinking monomers, or by first free radical polymerization of the sulfonyl zwitterionic monomers themselves and then block polymerization with crosslinking monomers. The slightly acid-responsive intelligent antibacterial surface of the present invention achieves intelligent antibacterial properties on the material surface by introducing a sulfonyl zwitterionic polymer modification layer onto the surface of the biomaterial and utilizing the charge interconversion of the modification layer under slightly acidic conditions. The slightly acid-responsive intelligent antibacterial surface of the present invention can effectively reduce bacterial adhesion on the material surface; when a small number of bacteria adhere to the surface, they are killed by the cationic groups in the modification layer after the surface charge is converted, and then the killed bacteria are released when the zwitterionic surface is restored.

Description

Technical Field

[0001] The present invention belongs to the technical field of polymer antibacterial materials, and in particular relates to a sulfonyl zwitterionic polymer with antibacterial function, a preparation method and an application thereof. Background Art

[0002] In modern society, bacterial infections associated with biomedical materials and medical devices have become a serious threat to human health. Bacteria attach, colonize, and proliferate on the surfaces of biomaterials, rapidly spreading through various surfaces, healthcare workers, and patients. Eradication is difficult, posing a significant challenge to healthcare. Currently, infections caused by implants account for over half of all hospital-acquired infections worldwide, afflicting a significant number of patients annually and resulting in significant medical costs.

[0003] In recent years, surfaces with antibacterial properties have provided an effective strategy for preventing bacterial infection of materials. Although these modified surfaces each have certain advantages, they still have problems such as complex preparation process, loading of antibacterial agents and high cytotoxicity. For example, 1. Some antibacterial surfaces loaded with antibiotics have bactericidal capabilities and can reduce the risk of infection, but the deposition of dead bacteria on the surface of the material will reduce the efficiency of further antibacterial treatment; and the use of antibiotics will aggravate the development of bacterial resistance. 2. Some antibacterial adhesion surfaces have antibacterial adhesion effects and prevent bacterial colonization and reproduction; but they cannot kill bacteria, and a small amount of bacteria attached to the surface will form a biofilm. 3. Some intelligent responsive antibacterial materials that use light, heat, salt and other conditions for stimulation can achieve the purpose of intelligent antibacterial, but they require additional stimulation from the external environment, which is inconvenient to use, and their preparation process is also relatively cumbersome and costly.

[0004] Therefore, developing and preparing an intelligent antibacterial material that can respond to the acidic microenvironment of bacteria themselves and solve related infection problems caused by bacteria is of great significance to human public health and the prevention of disease. Summary of the Invention

[0005] The technical purpose of the present invention is to address the technical problems of the prior art, such as the complex preparation process of antibacterial surfaces, the loading of antibacterial agents, and high cytotoxicity, by providing a sulfonyl zwitterionic monomer, a sulfonyl zwitterionic polymer, and a slightly acid-responsive intelligent antibacterial surface prepared using the sulfonyl zwitterionic polymer with antibacterial function. The slightly acid-responsive intelligent antibacterial surface introduces a sulfonyl zwitterionic polymer modification layer on the surface of the biomaterial and utilizes the charge interconversion of the modification layer under slightly acidic conditions to achieve the process of intelligent antibacterial on the material surface. The slightly acid-responsive intelligent antibacterial surface of the present invention can effectively reduce the adhesion of bacteria on the material surface; when a small amount of bacteria adhere to the surface, they are killed by the cationic groups after the surface charge of the modification layer is converted, and then the killed bacteria are released when the zwitterionic surface is restored.

[0006] In order to solve the above technical problems, the present invention adopts the following technical solution: a sulfonyl zwitterionic monomer having the structural formula:

[0007] The value range of q is 1-10.

[0008] Sulfonyl zwitterionic polymers with antibacterial function are prepared by polymerizing sulfonyl zwitterionic monomers with crosslinking monomers, or by first free radical polymerization of the sulfonyl zwitterionic monomers themselves and then block polymerization with the crosslinking monomers. The structural formula is as follows:

[0009] The value range of m is 1-90, and the value range of n is 1-30. The structure of A is:

[0010] And the value range of q in the formula is 1-10;

[0011] B is any one of isocyanoethyl methacrylate, N-(3-aminopropyl)-2-methyl-prop-2-enamide, N-(2-aminoethyl)methacrylamide hydrochloride, 3-methacrylamidophenylboronic acid, dopamine methacrylamide, dopamine acrylamide, 2-aminoethyl methacrylate hydrochloride, 2-aminoethyl methacrylate, hydroxyethyl methacrylate, hydroxyethyl acrylate, glycidyl methacrylate and γ-methacryloxypropyltrimethoxysilane.

[0012] Application of sulfonyl zwitterionic polymers in the preparation of slightly acid-responsive smart antibacterial surfaces.

[0013] The invention discloses a method for using a sulfonyl zwitterionic polymer to prepare a slightly acid-responsive intelligent antibacterial surface. The sulfonyl zwitterionic polymer is dissolved in a solvent and coated on the surface of a pre-modified medical material. The polymer is then cured at 25 to 120°C to obtain a slightly acid-responsive intelligent antibacterial surface.

[0014] Furthermore, the solvent is any one of methanol, ethanol and water, and the mass concentration of the sulfonyl zwitterionic polymer in the solvent is 5-30%; the coating method is any one of spin coating, dip coating and drop coating.

[0015] Furthermore, the modification method of the medical material is to place the clean medical material in a silane coupling agent solution for silanization treatment, and the silane coupling agent is any one of 3-aminopropyltriethoxysilane, N-(2-aminoethyl)-3-aminopropyltrimethoxysilane, γ-methacryloyloxypropyltrimethoxysilane, 3-(methacryloyloxy)propyltrimethoxysilane, 3-isocyanatepropyltrimethoxysilane and isocyanatepropyltriethoxysilane.

[0016] The preparation method of the sulfonyl zwitterionic monomer comprises the following steps:

[0017] 1) Weigh compound C, triethylamine and compound D in a molar ratio of 1:(1-3):(1-3), dissolve the weighed compound C and triethylamine in a dichloromethane solution, and stir in an ice-water bath for 10-30 minutes to prepare a mixed solution C. Then, dissolve the weighed compound D in a dichloromethane solution to prepare a mixed solution D. Then, under continuous stirring, add the prepared mixed solution D dropwise to the mixed solution C, and stir the reaction at a temperature of 25-37° C. for 12-24 hours. The obtained reaction product is filtered, washed, purified by column chromatography and dried to obtain a double-bond-terminated bromoester for later use.

[0018] Wherein, compound C is wherein r ranges from 1 to 10, and compound D is methacryloyl chloride or acryloyl chloride;

[0019] 2) dissolving 4-hydroxybenzenesulfonamide, tetrachlorobutyric acid, 1-hydroxybenzotriazole, 4-dimethylaminopyridine and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride in a dichloromethane solution at a molar ratio of 1:(1.5-3):(1.5-3):(2-3.5):(2-3.5), respectively, and carrying out a condensation reaction at 25-37° C. for 24-72 hours. The obtained reaction product is extracted, washed, purified by column chromatography and dried to obtain (4-chlorobutyryl)((4-hydroxyphenyl)sulfonyl)amide, which is then used for standby use;

[0020] 3) Dissolving the (4-chlorobutyryl)((4-hydroxyphenyl)sulfonyl)amide prepared in step 2) and the double-bond-terminated bromoester prepared in step 1) in an N,N-dimethylformamide solution in a molar ratio of 1:(1-3) respectively, then adding potassium carbonate in an amount of 1.5 to 4 times the molar amount of (4-chlorobutyryl)((4-hydroxyphenyl)sulfonyl)amide added thereto to provide an alkaline environment, and stirring the reaction at room temperature for 24-72 hours. The obtained reaction product is washed, centrifuged, and dried to obtain 2-(4-(N-(4-chlorobutyryl)sulfamoyl)phenoxy)ethyl methacrylate, which is then set aside;

[0021] 4) In a molar ratio of 1:(1-10), 2-(4-(N-(4-chlorobutyryl)sulfamoyl)phenoxy)ethyl methacrylate prepared in step 3) is placed in an aqueous trimethylamine solution and stirred at 65-80° C. for 12-72 hours. The reaction product is then filtered, concentrated, and freeze-dried to obtain a sulfonyl zwitterionic monomer.

[0022] The preparation method of a sulfonyl zwitterionic polymer with antibacterial function comprises the following steps: performing a polymerization reaction on a sulfonyl zwitterionic monomer and a crosslinking monomer, or first performing a self-radical polymerization reaction on the sulfonyl zwitterionic monomer and then performing a block polymerization reaction with the crosslinking monomer; the obtained reaction product is purified by dialysis and freeze-dried to obtain a finished sulfonyl zwitterionic polymer;

[0023] During the polymerization reaction, the reaction raw materials need to be dissolved in an organic solvent together with the initiator. The polymerization temperature is 65-75°C and the reaction time is 24-48 hours. When the cross-linking monomer undergoes polymerization, the molar ratio between it and the reaction material to be polymerized is 1: (2-10).

[0024] Furthermore, the initiator is AIBN or ACVA, and the added amount of the initiator is 1 / 100-1 / 1000 of the molar amount of the sulfonyl zwitterionic monomer.

[0025] Furthermore, the cross-linking monomer is any one of isocyanoethyl methacrylate, N-(3-aminopropyl)-2-methyl-prop-2-enamide, N-(2-aminoethyl) methacrylamide hydrochloride, 3-methacrylamidophenylboronic acid, dopamine methacrylamide, dopamine acrylamide, 2-aminoethyl methacrylate hydrochloride, 2-aminoethyl methacrylate, hydroxyethyl methacrylate, hydroxyethyl acrylate, glycidyl methacrylate and γ-methacryloxypropyltrimethoxysilane.

[0026] The beneficial effects of the present invention are:

[0027] 1. In the sulfonyl zwitterionic polymer structure designed and synthesized by the present invention, the anion can be protonated in a slightly acidic environment, and the cationic quaternary ammonium salt group can kill bacteria by contact. Then, after the microenvironment is restored, the zwitterionic surface is restored, and the killed bacteria are released.

[0028] 2. The slightly acid-responsive intelligent antibacterial surface prepared by the present invention utilizes the slightly acidic environment generated by the bacteria's own metabolism as the response condition. The modified layer itself does not need to be loaded with a bactericide, and can achieve the triple functions of anti-fouling, sterilization, and release, with good use effect and high safety and reliability.

[0029] 3. The sulfonyl zwitterionic polymer surface designed and prepared by the present invention has excellent anti-bacterial adhesion ability and blood compatibility; the surface bactericidal property of the sulfonyl zwitterionic polymer modified layer mainly depends on charge reversal, has no bactericide loading, and can avoid the development of bacterial resistance; in addition, compared with existing pH-responsive surfaces, this intelligent antibacterial functional surface responds in a slightly acidic environment of bacterial infection, does not require the stimulation of the external environment, is easy to use, low cost, and can automatically realize the intelligent antibacterial process. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 This is a diagram of the antibacterial process of the slightly acid-responsive intelligent antibacterial surface prepared by the present invention;

[0031] Figure 2 This is the H NMR spectrum of the sulfonyl zwitterionic monomer prepared in Example 1 of the present invention;

[0032] Figure 3 This is a comparison chart of the antibacterial performance of the slightly acid-responsive intelligent antibacterial surface prepared in Example 1 of the present invention under different environments;

[0033] Figure 4 This is a comparison photo of the antibacterial performance test of the slightly acid-responsive intelligent antibacterial surface prepared in Example 1 of the present invention against Escherichia coli and Staphylococcus aureus;

[0034] Figure 5 This is a diagram showing the in vitro biocompatibility test results of the slightly acid-responsive intelligent antibacterial surface prepared in Example 1 of the present invention. DETAILED DESCRIPTION

[0035] The embodiments of the present invention are described in detail in conjunction with specific implementation methods. This embodiment is based on the technical solution of the present invention and provides a detailed implementation method and specific operation process, but the protection scope of the present invention is not limited to the following embodiments.

[0036] The present invention provides a sulfonyl zwitterionic monomer, a sulfonyl zwitterionic polymer prepared using the sulfonyl zwitterionic monomer, and a slightly acid-responsive intelligent antibacterial surface prepared using the sulfonyl zwitterionic polymer.

[0037] The present invention's preparation process uses a sulfonyl zwitterionic polymer as a modified layer to create a surface with a slightly acid-responsive intelligent antibacterial function. This surface utilizes the slight acidification of the bacterial infection site as a response condition, achieving an intelligent antibacterial process combining the material's surface functions of "anti-adhesion, sterilization, and release."

[0038] The structural formula of the sulfonyl zwitterionic monomer is:

[0039] The value range of q is 1-10.

[0040] Amide zwitterionic polymers with antibacterial function, wherein the sulfonyl zwitterionic polymers are formed by polymerization of sulfonyl zwitterionic monomers and crosslinking monomers, or by free radical polymerization of the sulfonyl zwitterionic monomers themselves and then block polymerization with crosslinking monomers. The structural formula is as follows:

[0041] The value range of m is 1-90, and the value range of n is 1-30. The structure of A is:

[0042] And the value range of q in the formula is 1-10;

[0043] B is any one of isocyanoethyl methacrylate, N-(3-aminopropyl)-2-methyl-prop-2-enamide, N-(2-aminoethyl)methacrylamide hydrochloride, 3-methacrylamidophenylboronic acid, dopamine methacrylamide, dopamine acrylamide, 2-aminoethyl methacrylate hydrochloride, 2-aminoethyl methacrylate, hydroxyethyl methacrylate, hydroxyethyl acrylate, glycidyl methacrylate and γ-methacryloxypropyltrimethoxysilane.

[0044] A method for using a sulfonyl zwitterionic polymer to prepare a slightly acid-responsive intelligent antimicrobial surface comprises dissolving the sulfonyl zwitterionic polymer in a solvent and coating it on a pre-modified medical material surface. The polymer is then cured at 25-120°C to produce the slightly acid-responsive intelligent antimicrobial surface. The solvent is any one of methanol, ethanol, and water, and the mass concentration of the sulfonyl zwitterionic polymer in the solvent is 5-30%. The coating method is any one of spin coating, dip coating, and drop coating. The modification method of the medical material is to place the clean medical material in a silane coupling agent solution for silanization treatment, and the silane coupling agent is any one of 3-aminopropyltriethoxysilane, N-(2-aminoethyl)-3-aminopropyltrimethoxysilane, γ-methacryloyloxypropyltrimethoxysilane, 3-(methacryloyloxy)propyltrimethoxysilane, 3-isocyanatepropyltrimethoxysilane and isocyanatepropyltriethoxysilane.

[0045] The preparation method of the sulfonyl zwitterionic monomer comprises the following steps:

[0046] 1) Weigh compound C, triethylamine and compound D in a molar ratio of 1:(1-3):(1-3), dissolve the weighed compound C and triethylamine in a dichloromethane solution, and stir in an ice-water bath for 10-30 minutes to prepare a mixed solution C. Then, dissolve the weighed compound D in a dichloromethane solution to prepare a mixed solution D. Then, under continuous stirring, add the prepared mixed solution D dropwise to the mixed solution C, and stir the reaction at a temperature of 25-37° C. for 12-24 hours. The obtained reaction product is filtered, washed, purified by column chromatography and dried to obtain a double-bond-terminated bromoester for later use.

[0047] Wherein, compound C is wherein r ranges from 1 to 10, and compound D is methacryloyl chloride or acryloyl chloride;

[0048] 2) dissolving 4-hydroxybenzenesulfonamide, tetrachlorobutyric acid, 1-hydroxybenzotriazole, 4-dimethylaminopyridine and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride in a dichloromethane solution at a molar ratio of 1:(1.5-3):(1.5-3):(2-3.5):(2-3.5), respectively, and carrying out a condensation reaction at 25-37° C. for 24-72 hours. The obtained reaction product is extracted, washed, purified by column chromatography and dried to obtain (4-chlorobutyryl)((4-hydroxyphenyl)sulfonyl)amide, which is then used for standby use;

[0049] 3) Dissolving the (4-chlorobutyryl)((4-hydroxyphenyl)sulfonyl)amide prepared in step 2) and the double-bond-terminated bromoester prepared in step 1) in an N,N-dimethylformamide solution in a molar ratio of 1:(1-3) respectively, then adding potassium carbonate in an amount of 1.5 to 4 times the molar amount of (4-chlorobutyryl)((4-hydroxyphenyl)sulfonyl)amide added thereto to provide an alkaline environment, and stirring the reaction at room temperature for 24-72 hours. The obtained reaction product is washed, centrifuged, and dried to obtain 2-(4-(N-(4-chlorobutyryl)sulfamoyl)phenoxy)ethyl methacrylate, which is then set aside;

[0050] 4) In a molar ratio of 1:(1-10), 2-(4-(N-(4-chlorobutyryl)sulfamoyl)phenoxy)ethyl methacrylate prepared in step 3) is placed in an aqueous trimethylamine solution and stirred at 65-80° C. for 12-72 hours. The reaction product is then filtered, concentrated, and freeze-dried to obtain a sulfonyl zwitterionic monomer.

[0051] The preparation method of a sulfonyl zwitterionic polymer with antibacterial function comprises the following steps:

[0052] A sulfonyl zwitterionic monomer is subjected to a polymerization reaction with a crosslinking monomer, or a sulfonyl zwitterionic monomer is first subjected to a self-radical polymerization reaction and then to a block polymerization reaction with a crosslinking monomer. The resulting reaction product is purified by dialysis and freeze-dried to obtain a finished sulfonyl zwitterionic polymer;

[0053] During the polymerization reaction, the reaction raw materials need to be dissolved in an organic solvent together with the initiator. The polymerization temperature is 65-75°C and the reaction time is 24-48 hours. When the cross-linking monomer undergoes polymerization, the molar ratio between it and the reaction material to be polymerized is 1: (2-10).

[0054] The initiator is AIBN or ACVA, and the added amount of the initiator is 1 / 100-1 / 1000 of the molar added amount of the sulfonyl zwitterionic monomer.

[0055] The cross-linking monomer is any one of isocyanoethyl methacrylate, N-(3-aminopropyl)-2-methyl-prop-2-enamide, N-(2-aminoethyl) methacrylamide hydrochloride, 3-methacrylamidophenylboronic acid, dopamine methacrylamide, dopamine acrylamide, 2-aminoethyl methacrylate hydrochloride, 2-aminoethyl methacrylate, hydroxyethyl methacrylate, hydroxyethyl acrylate, glycidyl methacrylate and γ-methacryloyloxypropyltrimethoxysilane.

[0056] Example 1:

[0057] This embodiment provides a method for preparing a slightly acid-responsive intelligent antibacterial surface, which specifically includes the following steps:

[0058] (1) Preparation of sulfonyl zwitterionic monomers

[0059] First, 6.25 g of 2-bromoethanol (0.05 mol) and 8.3 mL of triethylamine (TEA) (0.06 mol) were dissolved in 7 mL of dichloromethane (DCM) solution and stirred under an ice-water bath. 5.83 g of methacryloyl chloride (0.06 mol) was mixed with 10 mL of DCM solution and added dropwise to the above mixed solution. The mixture was then stirred at room temperature overnight to obtain monomer 2-bromoethyl methacrylate.

[0060] 0.54 g of 4-hydroxybenzenesulfonamide, 0.57 g of tetrachlorobutyric acid, 0.42 g of 1-hydroxybenzotriazole (HOBt), 1.51 g of 4-dimethylaminopyridine (DMAP), and 2.3 g of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC·HCl) were dissolved in 60 mL of DCM and stirred at room temperature for 48 hours to obtain (4-chlorobutyryl)((4-hydroxyphenyl)sulfonyl)amide. Then, 194 mg of (4-chlorobutyryl)((4-hydroxyphenyl)sulfonyl)amide and 202.65 mg of 2-bromoethyl methacrylate were dissolved in 1 mL of DMF, and 121 mg of potassium carbonate was added. The mixture was stirred at room temperature for 72 hours to obtain 2-(4-(N-(4-chlorobutyryl)sulfamoyl)phenoxy)ethyl methacrylate.

[0061] Finally, 2-(4-(N-(4-chlorobutyryl)sulfamoyl)phenoxy)ethyl methacrylate was stirred in 5 mL of trimethylamine aqueous solution (TMA / H2O) at 70°C for 24 hours, and the reaction mixture was filtered, concentrated and freeze-dried to obtain ((4-(2-(methacryloyloxy)ethoxy)phenyl)sulfonyl)(4-(trimethylamino)butyryl)amide monomer (MPTA).

[0062]

[0063] Synthesis process of monomer 2-bromoethyl methacrylate

[0064]

[0065] The synthesis process of monomer MPTA uses deuterated chloroform as solvent. The nuclear magnetic hydrogen spectrum of the MPTA zwitterionic monomer prepared in this example is shown in the attached figure. Figure 2 shown.

[0066] (2) Preparation of zwitterionic copolymer P(MPTA-AEMA)

[0067] In this example, the polymer was prepared using conventional free radical polymerization. 230.8 mg of the zwitterionic monomer MPTA (0.56 mmol) prepared in step (1) and 39.4 mg of AEMA (0.24 mmol) were dissolved in 1.5 mL of DMF. 2.24 mg of ACVA (initiator) was added, and polymerization was carried out at 70°C for 24 h. After dialysis, the product was dehydrogenated and acidified in a weakly alkaline solution, followed by freeze-drying to obtain the polymer P(MPTA-AEMA).

[0068]

[0069] Synthesis process of zwitterionic copolymer P(MPTA-AEMA)

[0070] (3) Preparation of slightly acid-responsive intelligent antibacterial surface

[0071] The medical gauze was first ultrasonically treated in an ethanol solution for 30 minutes and dried for later use. The pretreated medical gauze was then placed in a 5% γ-glycidyloxypropyltrimethoxysilane (KH560) solution and subjected to silanization treatment under reflux for 24 hours. The gauze was then removed and the residual KH560 on the surface was cleaned with an ethanol solution. The gauze was then dried for later use. The KH560-treated medical gauze was dipped in a 15% zwitterionic copolymer P(MPTA-AEMA) solution prepared in step (2) for 20 minutes. The gauze was then removed and cured at 80°C to obtain a finished product, the P(MPTA-AEMA) type antibacterial surface (PMA).

[0072] Example 2:

[0073] This embodiment provides a method for preparing a slightly acid-responsive intelligent antibacterial surface, which specifically includes the following steps:

[0074] (1) Preparation of sulfonyl zwitterionic monomers

[0075] First, 6.25 g of 2-bromoethanol (0.05 mol) and 8.3 mL of triethylamine (TEA) (0.06 mol) were dissolved in 7 mL of dichloromethane (DCM) solution and stirred under an ice-water bath. 5.43 g of acryloyl chloride (0.06 mol) was mixed with 10 mL of DCM solution and added dropwise to the above mixed solution. The mixture was then stirred at room temperature overnight to obtain monomer 2-bromoethyl acrylate.

[0076] 0.54 g of 4-hydroxybenzenesulfonamide, 0.57 g of tetrachlorobutyric acid, 0.42 g of 1-hydroxybenzotriazole (HOBt), 1.51 g of 4-dimethylaminopyridine (DMAP), and 2.3 g of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC·HCl) were dissolved in 60 mL of DCM and stirred at room temperature for 48 hours to obtain (4-chlorobutyryl)((4-hydroxyphenyl)sulfonyl)amide. Then, 194 mg of (4-chlorobutyryl)((4-hydroxyphenyl)sulfonyl)amide and 187.92 mg of 2-bromoethyl acrylate were dissolved in 1 mL of DMF, and 121 mg of potassium carbonate was added. The mixture was stirred at room temperature for 72 hours to obtain 2-(4-(N-(4-chlorobutyryl)sulfamoyl)phenoxy)ethyl acrylate.

[0077] Finally, 2-(4-(N-(4-chlorobutyryl)sulfamoyl)phenoxy)ethyl acrylate was stirred in 5 mL of trimethylamine aqueous solution (TMA / H2O) at 70°C for 24 hours, and the reaction mixture was filtered, concentrated and freeze-dried to obtain ((4-(2-(acryloyloxy)ethoxy)phenyl)sulfonyl)(4-(trimethylamino)butyryl)amide monomer (APTA).

[0078]

[0079] Synthesis process of monomer 2-bromoethyl acrylate

[0080]

[0081] Synthesis process of monomer APTA

[0082] (2) Preparation of zwitterionic polymer P(APTA-co-AEMA)

[0083] In this example, a copolymer was prepared by RAFT polymerization. 223.14 mg of the zwitterionic monomer APTA (0.56 mmol) obtained in step (1) and 39.4 mg of N-(2-aminoethyl) methacrylamide hydrochloride (AEMA) (0.24 mmol) were dissolved in 1.5 mL of N,N-dimethylformamide (DMF) solution. A chain initiator, 4.47 mg of 4-cyano-4-(thiobenzoyl) valeric acid, and an initiator, 1.12 mg of ACVA, were added. The polymerization reaction was carried out at 70° C. for 24 h. After dialysis, the mixture was dehydrogenated and acidified in a weak alkaline solution, and then freeze-dried to obtain a polymer P(APTA-co-AEMA).

[0084]

[0085] Synthesis process of zwitterionic polymer P(APTA-co-AEMA)

[0086] (3) Preparation of slightly acid-responsive intelligent antibacterial surface

[0087] First, the medical gauze is ultrasonically treated in an ethanol solution for 30 minutes and dried for later use. The pretreated medical gauze is then placed in an 8% N-(2-aminoethyl)-3-aminopropyltrimethoxysilane solution and subjected to silanization treatment under reflux for 24 hours. The gauze is then removed and the residual silane coupling agent on the surface is removed with a methanol solution. The gauze is then dried for later use. The silanized medical gauze is then dipped in a 20% zwitterionic copolymer P(APTA-co-AEMA) solution prepared in step (2) for 15 minutes. The gauze is then removed and cured at 70°C to obtain a finished P(APTA-co-AEMA) antibacterial surface.

[0088] Example 3:

[0089] This embodiment provides a method for preparing a slightly acid-responsive intelligent antibacterial surface, which specifically includes the following steps:

[0090] (1) Preparation of sulfonyl zwitterionic monomers

[0091] First, 6.95 g of 3-bromopropanol (0.05 mol) and 8.3 mL of triethylamine (TEA) (0.06 mol) were dissolved in 7 mL of dichloromethane (DCM) solution and stirred under an ice-water bath. 5.83 g of methacryloyl chloride (0.06 mol) was mixed with 10 mL of DCM solution and added dropwise to the above mixed solution. The mixture was then stirred at room temperature overnight to obtain the monomer 3-bromopropyl methacrylate.

[0092] 0.54 g of 4-hydroxybenzenesulfonamide, 0.57 g of tetrachlorobutyric acid, 0.42 g of 1-hydroxybenzotriazole (HOBt), 1.51 g of 4-dimethylaminopyridine (DMAP), and 2.3 g of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC·HCl) were dissolved in 60 mL of DCM and stirred at room temperature for 48 hours to obtain (4-chlorobutyryl)((4-hydroxyphenyl)sulfonyl)amide. Subsequently, 194 mg of (4-chlorobutyryl)((4-hydroxyphenyl)sulfonyl)amide and 217.38 mg of 3-bromopropyl methacrylate were dissolved in 1 mL of DMF, and 121 mg of potassium carbonate was added. The mixture was stirred at room temperature for 72 hours to obtain ((4-(3-(methacryloyloxy)propoxy)phenyl)sulfonyl)(4-chlorobutyryl)amide.

[0093] Finally, ((4-(3-(methacryloyloxy)propoxy)phenyl)sulfonyl)(4-chlorobutyryl)amide was stirred in 5 mL of trimethylamine aqueous solution (TMA / H2O) at 70°C for 24 hours, and the reaction mixture was filtered, concentrated and freeze-dried to obtain ((4-(3-(methacryloyloxy)propoxy)phenyl)sulfonyl)(4-(trimethylamino)butyryl)amide monomer (MPPTA).

[0094]

[0095] Synthesis process of monomer 3-bromopropyl methacrylate

[0096]

[0097] Synthesis process of monomer MPPTA

[0098] (2) Preparation of zwitterionic block polymer P(MPPTA-b-AEMA)

[0099] 213.27 mg of the MPPTA amphiphilic monomer prepared in step (1) was dissolved in 1 mL of DMF solution, and a chain initiator, 2.0 mg of 4-cyano-4-(thiobenzoyl)valeric acid, and an initiator, 0.67 mg of ACVA, were added. The polymerization reaction was carried out at 70°C for 24 hours, followed by dialysis and freeze-drying to obtain PMPPTA. PMPPTA and AEMA were then dissolved in DMF solution, and the initiator, ACVA, was added. The reaction was carried out at 70°C for 24 hours. After dialysis, the product was dehydrogenated and treated in a weak alkaline solution, and then freeze-dried to obtain a block polymer, P(MPPTA-b-AEMA).

[0100]

[0101] Synthesis process of zwitterionic block polymer P(MPPTA-b-AEMA)

[0102] (3) Preparation of slightly acid-responsive intelligent antibacterial surface

[0103] The medical gauze is first ultrasonically treated in a methanol solution for 30 minutes and dried for later use. The pretreated medical gauze is then placed in a 6% isocyanatepropyltriethoxysilane solution and subjected to silanization treatment under reflux conditions for 24 hours. The gauze is then removed and the residual silane coupling agent on the surface is removed with a methanol solution. The gauze is then dried for later use. The 30% zwitterionic copolymer P (MPPTA-b-AEMA) solution prepared in step (2) is repeatedly spin-coated on the surface of the silanized medical gauze by spin coating. The gauze is then cured at 120° C. to obtain a finished P (MPPTA-b-AEMA) antibacterial surface.

[0104] Example 4:

[0105] This embodiment provides a method for preparing a slightly acid-responsive intelligent antibacterial surface, which specifically includes the following steps:

[0106] (1) Preparation of sulfonyl zwitterionic monomers

[0107] The specific operation of this step is the same as that of Example 1.

[0108] (2) Preparation of zwitterionic copolymer P(MPTA-GMA)

[0109] In this example, the polymer was prepared using conventional free radical polymerization. 288.75 mg of the zwitterionic monomer MPTA (0.7 mmol) prepared in step (1) and 42.65 mg of glycidyl methacrylate (GMA) (0.3 mmol) were dissolved in 2 mL of DMF. 2.80 mg of ACVA (initiator) was added, and polymerization was carried out at 70°C for 24 h. After dialysis, the product was dehydrogenated and acidified in a weakly alkaline solution, followed by freeze-drying to obtain polymer P(MPTA-GMA).

[0110] (3) Preparation of slightly acid-responsive intelligent antibacterial surface

[0111] The specific operation of this step is the same as that of Example 1, and the finished product P (MPTA-GMA) type antibacterial surface is finally obtained.

[0112] Example 5:

[0113] This embodiment provides a method for preparing a slightly acid-responsive intelligent antibacterial surface, which specifically includes the following steps:

[0114] (1) Preparation of sulfonyl zwitterionic monomers

[0115] The specific operation of this step is the same as that of Example 1.

[0116] (2) Preparation of zwitterionic copolymer P(MPTA-HEMA)

[0117] In this example, the polymer was prepared using conventional free radical polymerization. 288.75 mg of the MPTA (0.7 mmol) zwitterionic monomer prepared in step (1) and 39.04 mg of hydroxyethyl methacrylate (HEMA) (0.3 mmol) were dissolved in 2 mL of DMF. 2.80 mg of ACVA (initiator) was added, and polymerization was carried out at 70°C for 24 h. After dialysis, the product was dehydrogenated and acidified in a weakly alkaline solution, followed by freeze-drying to obtain the polymer P(MPTA-HEMA).

[0118] (3) Preparation of slightly acid-responsive intelligent antibacterial surface

[0119] The specific operation of this step is the same as that of Example 1, and the finished product P (MPTA-HEMA) type antibacterial surface is finally obtained.

[0120] Example 6:

[0121] This embodiment provides a method for preparing a slightly acid-responsive intelligent antibacterial surface, which specifically includes the following steps:

[0122] (1) Preparation of sulfonyl zwitterionic monomers

[0123] The specific operation of this step is the same as that of Example 1.

[0124] (2) Preparation of zwitterionic copolymer P(MPTA-KH570)

[0125] In this example, a polymer was prepared using conventional free radical polymerization. 288.75 mg of the MPTA (0.7 mmol) zwitterionic monomer prepared in step (1) and 74.51 mg of γ-methacryloxypropyltrimethoxysilane (KH570) (0.3 mmol) were dissolved in 2 mL of DMF. 2.80 mg of ACVA (initiator) was added, and polymerization was carried out at 70°C for 24 h. After dialysis, the product was dehydrogenated and acidified in a weakly alkaline solution, followed by freeze-drying to obtain polymer P(MPTA-KH570).

[0126] (3) Preparation of slightly acid-responsive intelligent antibacterial surface

[0127] The specific operation of this step is the same as that of Example 1, and the finished product P (MPTA-KH570) type antibacterial surface is finally obtained.

[0128] Example 7:

[0129] This embodiment provides a method for preparing a slightly acid-responsive intelligent antibacterial surface, which specifically includes the following steps:

[0130] (1) Preparation of sulfonyl zwitterionic monomers

[0131] The specific operation of this step is the same as that of Example 1.

[0132] (2) Preparation of zwitterionic copolymer P(MPTA-DMA)

[0133] In this example, the polymer was prepared using conventional free radical polymerization. 288.75 mg of the MPTA (0.7 mmol) zwitterionic monomer prepared in step (1) and 66.38 mg of dopamine methacrylamide (DMA) (0.3 mmol) were dissolved in 2 mL of DMF. 2.80 mg of ACVA (initiator) was added, and polymerization was carried out at 70°C for 24 h. After dialysis, the product was dehydrogenated and acidified in a weakly alkaline solution, followed by freeze-drying to obtain polymer P(MPTA-DMA).

[0134] (3) Preparation of slightly acid-responsive intelligent antibacterial surface

[0135] The specific operation of this step is the same as that of Example 1, and the finished product P (MPTA-DMA) type antibacterial surface is finally obtained.

[0136] Example 8:

[0137] This embodiment provides a method for preparing a slightly acid-responsive intelligent antibacterial surface, which specifically includes the following steps:

[0138] (1) Preparation of sulfonyl zwitterionic monomers

[0139] The specific operation of this step is the same as that of Example 1.

[0140] (2) Preparation of zwitterionic copolymer P(MPTA-IEM)

[0141] In this example, the polymer was prepared using conventional free radical polymerization. 288.75 mg of the MPTA (0.7 mmol) zwitterionic monomer prepared in step (1) and 46.55 mg of isocyanoethyl methacrylate (IEM) (0.3 mmol) were dissolved in 2 mL of DMF. 2.80 mg of ACVA (initiator) was added, and polymerization was carried out at 70°C for 24 h. After dialysis, the product was dehydrogenated and acidified in a weakly alkaline solution, followed by freeze-drying to obtain polymer P(MPTA-IEM).

[0142] (3) Preparation of slightly acid-responsive intelligent antibacterial surface

[0143] The specific operation of this step is the same as that of Example 1, and the finished product P (MPTA-IEM) type antibacterial surface is finally obtained.

[0144] Experimental determination

[0145] 1. Surface micro-acid responsiveness

[0146] The antibacterial sample surface prepared in Example 1 of the present invention was placed in a physiological environment and a bacterial infection slightly acidic environment and co-cultured with an Escherichia coli suspension, and the surface after culture was subjected to live / dead staining to evaluate its slightly acidic responsiveness. Figure 3As shown, the surface is co-cultured with Escherichia coli, and the number of live and dead Escherichia coli bacteria is statistically analyzed after live / dead bacteria staining. The surface is covered with live bacteria in a neutral environment; while in a slightly acidic environment, the bacteria are killed by the quaternary ammonium groups on the surface after protonation, and only a small number of live bacteria exist, indicating that the surface has slightly acid responsiveness.

[0147] 2. Antibacterial properties

[0148] The plate colony count experiment was used to evaluate the antibacterial rate of the surface of the antibacterial sample prepared in Example 1 of the present invention against Escherichia coli and Staphylococcus aureus. Figure 4 As shown, the plate colony results after the surface was co-cultured with Escherichia coli and Staphylococcus aureus were respectively shown. After counting, the formula: R(%) = (BA) / B×100%, where R is the bacterial inhibition rate (%), A is the bacterial colony count result of the experimental group, and B is the bacterial colony result of the original substrate control group, was used to obtain the surface inhibition rate for Escherichia coli of 93.2%, and the inhibition rate for Staphylococcus aureus of 91.6%, indicating that the surface has good anti-bacterial adhesion ability.

[0149] 2. In vitro biocompatibility test

[0150] The MTT method was used to study the biocompatibility of the antibacterial surface prepared in Example 1 of the present invention with L929 cell line. Figure 5 As shown, the antimicrobial surface was co-cultured with cells for 24, 48, and 72 hours, respectively, and cell proliferation was measured using the MTT assay. The results showed that the relative cell growth rate on the antimicrobial surface was above 90% after each co-culture period. Based on the cytotoxicity grading criteria, the surface did not significantly affect cell growth.

[0151] It should be noted that although the present invention has been described with reference to the above embodiments, the present invention may also have other various embodiments. Without departing from the spirit and scope of the present invention, it is obvious that those skilled in the art may make various corresponding changes and modifications to the present invention, and such changes and modifications shall fall within the scope of protection of the appended claims and their equivalents.

Claims

1. A sulfonyl zwitterionic monomer, characterized in that: The structural formula is: The value range of q is 1-10.

2. Sulfonyl zwitterionic polymer with antibacterial function, characterized by: The sulfonyl zwitterionic polymer is formed by polymerizing a sulfonyl zwitterionic monomer and a crosslinking monomer, or by firstly free radical polymerization of the sulfonyl zwitterionic monomer itself and then block polymerization with the crosslinking monomer. The structural formula is as follows: The value range of m is 1-90, and the value range of n is 1-30. The structure of A is: And the value range of q in the formula is 1-10; B is any one of isocyanoethyl methacrylate, N-(3-aminopropyl)-2-methyl-prop-2-enamide, N-(2-aminoethyl)methacrylamide hydrochloride, 3-methacrylamidophenylboronic acid, dopamine methacrylamide, dopamine acrylamide, 2-aminoethyl methacrylate hydrochloride, 2-aminoethyl methacrylate, hydroxyethyl methacrylate, hydroxyethyl acrylate, glycidyl methacrylate and γ-methacryloxypropyltrimethoxysilane.

3. Use of the sulfonyl zwitterionic polymer with antibacterial function according to claim 2 in preparing a slightly acid-responsive intelligent antibacterial surface.

4. Use of the sulfonyl zwitterionic polymer according to claim 3 in preparing a slightly acid-responsive intelligent antibacterial surface, characterized in that: The sulfonyl zwitterionic polymer is dissolved in a solvent and coated on the surface of a pre-modified medical material. It is then cured at 25-120°C to obtain a slightly acid-responsive intelligent antibacterial surface.

5. Use of the sulfonyl zwitterionic polymer according to claim 4 in preparing a slightly acid-responsive intelligent antibacterial surface, characterized in that: The solvent is any one of methanol, ethanol and water, and the mass concentration of the sulfonyl zwitterionic polymer in the solvent is 5-30%; the coating method is any one of spin coating, dip coating and drop coating.

6. Use of the sulfonyl zwitterionic polymer according to claim 4 in preparing a slightly acid-responsive intelligent antibacterial surface, characterized in that: The modification method of the medical material is to place the clean medical material in a silane coupling agent solution for silanization treatment, and the silane coupling agent is any one of 3-aminopropyltriethoxysilane, N-(2-aminoethyl)-3-aminopropyltrimethoxysilane, γ-methacryloyloxypropyltrimethoxysilane, 3-(methacryloyloxy)propyltrimethoxysilane, 3-isocyanatepropyltrimethoxysilane and isocyanatepropyltriethoxysilane.

7. The method for preparing a sulfonyl zwitterionic monomer according to claim 1, wherein: The following steps are involved: 1) Weigh compound C, triethylamine and compound D in a molar ratio of 1:(1-3):(1-3), dissolve the weighed compound C and triethylamine in a dichloromethane solution, and stir in an ice-water bath for 10-30 minutes to prepare a mixed solution C. Then, dissolve the weighed compound D in a dichloromethane solution to prepare a mixed solution D. Then, under continuous stirring, add the prepared mixed solution D dropwise to the mixed solution C, and stir the reaction at a temperature of 25-37° C. for 12-24 hours. The obtained reaction product is filtered, washed, purified by column chromatography and dried to obtain a double-bond-terminated bromoester for later use. Wherein, compound C is wherein r ranges from 1 to 10, and compound D is methacryloyl chloride or acryloyl chloride; 2) dissolving 4-hydroxybenzenesulfonamide, tetrachlorobutyric acid, 1-hydroxybenzotriazole, 4-dimethylaminopyridine and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride in a dichloromethane solution at a molar ratio of 1:(1.5-3):(1.5-3):(2-3.5):(2-3.5), respectively, and carrying out a condensation reaction at 25-37° C. for 24-72 hours. The obtained reaction product is extracted, washed, purified by column chromatography and dried to obtain (4-chlorobutyryl)((4-hydroxyphenyl)sulfonyl)amide, which is then used for standby use; 3) Dissolving the (4-chlorobutyryl)((4-hydroxyphenyl)sulfonyl)amide prepared in step 2) and the double-bond-terminated bromoester prepared in step 1) in an N,N-dimethylformamide solution in a molar ratio of 1:(1-3) respectively, then adding potassium carbonate in an amount of 1.5 to 4 times the molar amount of (4-chlorobutyryl)((4-hydroxyphenyl)sulfonyl)amide added thereto to provide an alkaline environment, and stirring the reaction at room temperature for 24-72 hours. The obtained reaction product is washed, centrifuged, and dried to obtain 2-(4-(N-(4-chlorobutyryl)sulfamoyl)phenoxy)ethyl methacrylate, which is then set aside; 4) In a molar ratio of 1:(1-10), 2-(4-(N-(4-chlorobutyryl)sulfamoyl)phenoxy)ethyl methacrylate prepared in step 3) is placed in an aqueous trimethylamine solution and stirred at 65-80° C. for 12-72 hours. The reaction product is then filtered, concentrated, and freeze-dried to obtain a sulfonyl zwitterionic monomer.

8. The method for preparing the sulfonyl zwitterionic polymer with antibacterial function according to claim 2, characterized in that: The following steps are involved: A sulfonyl zwitterionic monomer is subjected to a polymerization reaction with a crosslinking monomer, or a sulfonyl zwitterionic monomer is subjected to a free radical polymerization reaction by itself and then to a block polymerization reaction with a crosslinking monomer. The resulting reaction product is purified by dialysis and freeze-dried to obtain a finished sulfonyl zwitterionic polymer; During the polymerization reaction, the reaction raw materials need to be dissolved in an organic solvent together with the initiator. The polymerization temperature is 65-75°C and the reaction time is 24-48 hours. When the cross-linking monomer undergoes polymerization, the molar ratio between it and the reaction material to be polymerized is 1: (2-10).

9. The method for preparing the sulfonyl zwitterionic polymer with antibacterial function according to claim 8, characterized in that: The initiator is AIBN or ACVA, and the added amount of the initiator is 1 / 100-1 / 1000 of the molar added amount of the sulfonyl zwitterionic monomer.

10. The method for preparing the sulfonyl zwitterionic polymer with antibacterial function according to claim 8, characterized in that: The cross-linking monomer is any one of isocyanoethyl methacrylate, N-(3-aminopropyl)-2-methyl-prop-2-enamide, N-(2-aminoethyl) methacrylamide hydrochloride, 3-methacrylamidophenylboronic acid, dopamine methacrylamide, dopamine acrylamide, 2-aminoethyl methacrylate hydrochloride, 2-aminoethyl methacrylate, hydroxyethyl methacrylate, hydroxyethyl acrylate, glycidyl methacrylate and γ-methacryloyloxypropyltrimethoxysilane.

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