CN108659168B

Movatterモバイル変換

Info

Publication number: CN108659168B
Application number: CN201810539671.3A
Authority: CN
Inventors: 李树娜; 贾园; 杜威; 刘振; 朱刚; 方振华
Original assignee: Xian Unversity of Arts and Science
Current assignee: Xian Unversity of Arts and Science
Priority date: 2018-05-30
Filing date: 2018-05-30
Publication date: 2020-12-11
Anticipated expiration: 2038-05-30
Also published as: CN108659168A

Abstract

Translated fromChinese

本发明属于材料表面科学和生物医用高分子材料技术领域，具体涉及一种基于五氟苯基接枝反应合成的双仿生聚合物，并进一步公开其制备方法和应用。本发明所述双仿生聚合物以含有磷酰胆碱基团的乙烯基单体和含有五氟苯基的乙烯基单体为原料进行自由基聚合反应，得到含有五氟苯基的磷酰胆碱聚合物，并将仿蚌类粘附蛋白成分多巴胺与含有五氟苯基的磷酰胆碱聚合物相结合，邻二酚羟基具有仿蚌类粘附蛋白多巴胺的粘附作用，赋予水溶性聚合物在材料表面优异的粘附性能，磷酰胆碱具有仿细胞外层膜结构的抗污性能，获得了良好的抗生物污染的性能，该聚合物具有双重仿生的特征，在材料表面修饰领域具有广阔的应用前景。

The invention belongs to the technical fields of material surface science and biomedical polymer materials, in particular to a double biomimetic polymer synthesized based on a pentafluorophenyl graft reaction, and further discloses a preparation method and application thereof. The dual biomimetic polymer of the present invention uses a vinyl monomer containing a phosphorylcholine group and a vinyl monomer containing a pentafluorophenyl group as raw materials to carry out a free radical polymerization reaction to obtain a phosphorylcholine containing a pentafluorophenyl group Alkaline polymer, combining dopamine, a component of mussel-like adhesion proteins, with phosphorylcholine polymer containing pentafluorophenyl, and the ortho-diphenol hydroxyl group has the adhesion of mussel-like adhesion protein dopamine, giving water solubility The polymer has excellent adhesion performance on the surface of the material. Phosphorylcholine has anti-fouling performance that mimics the structure of the outer membrane of the cell, and obtains good anti-biological fouling performance. The polymer has dual biomimetic characteristics and is modified on the surface of the material. The field has broad application prospects.

Description

Translated fromChinese

一种双仿生聚合物及其制备方法与应用A kind of double biomimetic polymer and its preparation method and application

技术领域technical field

本发明属于材料表面科学和生物医用高分子材料技术领域，具体涉及一种基于五氟苯基接枝反应合成的双仿生聚合物，并进一步公开其制备方法和应用。The invention belongs to the technical fields of material surface science and biomedical polymer materials, in particular to a double biomimetic polymer synthesized based on a pentafluorophenyl graft reaction, and further discloses a preparation method and application thereof.

背景技术Background technique

生物相容性是指材料与生物体之间相互作用后产生的各种生物、物理、化学等反应的一种概念。一般地讲，就是材料植入人体后与人体相容程度，也就是说是否会对人体组织造成毒害作用。通常情况下，一般材料用于生物体内时易产生非特异性吸附蛋白，激活补体分子及免疫系统，从而引起凝血、免疫及炎症反应，以致其性能显著降低、甚至失效，这便是由于材料生物相容性较差的缘故。因而，生物相容性研究已成为生物材料研究领域中的首要问题。Biocompatibility refers to a concept of various biological, physical, chemical and other reactions generated by the interaction between materials and living organisms. Generally speaking, it is the degree of compatibility of the material with the human body after implantation into the human body, that is, whether it will cause toxic effects on human tissues. Under normal circumstances, when general materials are used in the body, they are prone to produce non-specific adsorption proteins, activate complement molecules and the immune system, thereby causing coagulation, immune and inflammatory responses, resulting in significantly reduced performance or even failure. This is due to the biological phase of the material. due to poor compatibility. Therefore, biocompatibility research has become the primary issue in the field of biomaterial research.

由于材料表面是材料与生物体接触的媒介，材料表面的电荷、亲/疏水性、化学组成、形貌等都是影响材料与生物体之间界面相互作用的重要因素，也是决定材料生物相容性是否优异的主要因素。因此，提高材料表面的生物相容性是解决这一科学问题的关键。而将具有良好生物相容性的物质引入到材料表面进行改性则是改善材料与生物体之间的相互作用、提高材料生物相容性最简便而有效的途径。因此，材料表面的生物相容性改性是生物材料研究领域中一个永恒的主题，具有重要的学术意义和巨大的应用前景。Since the surface of the material is the medium for the contact between the material and the organism, the charge, hydrophilicity/hydrophobicity, chemical composition, and morphology of the material surface are all important factors that affect the interface interaction between the material and the organism, and also determine the biocompatibility of the material. The main factor of whether the sex is excellent. Therefore, improving the biocompatibility of material surfaces is the key to solving this scientific problem. Introducing a substance with good biocompatibility into the surface of the material for modification is the most convenient and effective way to improve the interaction between the material and the organism and improve the biocompatibility of the material. Therefore, the biocompatibility modification of material surfaces is an eternal topic in the field of biomaterials research, with important academic significance and great application prospects.

已有报道表明，将具有良好血液相容性的内皮细胞、白蛋白、肝素和/或聚乙二醇引入到材料表面，可以明显改善其生物相容性，特别是显著提高其血液相容性。但是，上述用于改善材料表面生物相容性的改性材料中均存在着各种各样的问题，例如内皮细胞与材料表面的相互作用较差、抗血液冲击能力不佳、易脱落等问题；并且由于白蛋白与体内活性组分在材料表面竞争吸附，导致吸附在材料表面的白蛋白含量降低，甚至变性；而肝素易水解，致使其活性明显下降，以致诱导出血、血小板减少症等并发症；在猛烈呼吸过程中，聚乙二醇因超氧阴离子和过氧化氢而被氧化，其表面也有不同程度生物污染。可见，现有用于改善材料表面生物相容性的改性材料的性能并不理想。It has been reported that the introduction of endothelial cells, albumin, heparin and/or polyethylene glycol with good blood compatibility to the surface of the material can significantly improve its biocompatibility, especially its blood compatibility. . However, there are various problems in the above-mentioned modified materials for improving the biocompatibility of the material surface, such as poor interaction between endothelial cells and the material surface, poor resistance to blood shock, and easy shedding. ; And because albumin competes with the active components in the body for adsorption on the surface of the material, the content of albumin adsorbed on the surface of the material is reduced, or even denatured; and heparin is easily hydrolyzed, resulting in a significant decrease in its activity, resulting in the induction of bleeding, thrombocytopenia and other complications. Symptoms; in the process of vigorous breathing, polyethylene glycol is oxidized due to superoxide anion and hydrogen peroxide, and its surface is also biologically contaminated to varying degrees. It can be seen that the performance of the existing modified materials for improving the biocompatibility of the material surface is not ideal.

磷酰胆碱(phosphorylcholine，PC)是组成细胞膜基本单元卵磷脂的亲水端基，是细胞外层膜中的外层官能团，同时带有正、负异种电荷，具有较强的结合水的能力和亲水性能，这种结构和组成的表面与生理环境相互作用不仅不会吸附和沉积蛋白质，也不会引发血小板激活、导致凝血等不良反应，具有良好生物相容性。近几年来的研究表明，采用磷酰胆碱基团及其聚合物在材料表面构建具有仿细胞外层膜结构，可以显著改善材料的血液相容性。Phosphorylcholine (PC) is the hydrophilic end group of lecithin, the basic unit of cell membrane, and the outer functional group in the outer membrane of the cell. And hydrophilic properties, the surface of this structure and composition interacts with the physiological environment not only does not adsorb and deposit proteins, but also does not trigger platelet activation, lead to coagulation and other adverse reactions, and has good biocompatibility. Studies in recent years have shown that the use of phosphorylcholine groups and their polymers to construct a cell-like outer membrane structure on the surface of the material can significantly improve the blood compatibility of the material.

现有进行材料表面生物相容性改性的方法主要为物理涂覆，即包括浸涂、旋涂和滴涂等方式，并因其具有工艺简单、操作方便和条件温和的优点，是构建仿细胞外层膜结构获得优异生物相容性表面的理想手段。然而，对于磷酰胆碱改性材料而言，由于其基团的亲水性较强，使得通过物理涂覆在材料表面的磷酰胆碱聚合物涂层在复杂的生理环境中容易发生溶解、降解、甚至脱落。因而，需要将可交联或共价键基团引入到磷酰胆碱聚合物中，经过化学反应将该聚合物涂层交联或共价键合在材料表面。这无疑增加了这类磷酰胆碱聚合物合成及应用对表面要求的难度，也使得该技术的处理过程冗长复杂，因此，迫切需要研究开发使用简单、适用面广的表面改性方法。The existing methods for modifying the surface biocompatibility of materials are mainly physical coating, including dip coating, spin coating and drop coating. An ideal means of obtaining an excellent biocompatible surface for the structure of the outer membrane of the cell. However, for phosphorylcholine-modified materials, due to the strong hydrophilicity of their groups, the phosphorylcholine polymer coating that is physically coated on the surface of the material is easily dissolved in a complex physiological environment. , degradation, or even fall off. Therefore, it is necessary to introduce crosslinkable or covalently bonded groups into the phosphorylcholine polymer, and to crosslink or covalently bond the polymer coating to the surface of the material through a chemical reaction. This undoubtedly increases the difficulty of surface requirements for the synthesis and application of this type of phosphorylcholine polymer, and also makes the treatment process of this technology long and complicated. Therefore, it is urgent to research and develop a simple and widely applicable surface modification method.

发明内容SUMMARY OF THE INVENTION

为此，本发明所要解决的技术问题在于提供一种双仿生聚合物，以解决现有技术中磷酰胆碱聚合物进行材料表面生物相容性改性时涂层容易发生溶解、降解、甚至脱落的问题；Therefore, the technical problem to be solved by the present invention is to provide a dual biomimetic polymer, so as to solve the problem that when the phosphorylcholine polymer in the prior art is modified for the biocompatibility of the material surface, the coating is easily dissolved, degraded, or even the problem of falling off;

本发明解决的第二个技术问题在于提供上述双仿生聚合物的制备方法。The second technical problem solved by the present invention is to provide a method for preparing the above-mentioned dual biomimetic polymer.

为解决上述技术问题，本发明所述的一种双仿生聚合物的制备方法，包括如下步骤：In order to solve the above-mentioned technical problems, the preparation method of a dual biomimetic polymer of the present invention comprises the following steps:

(1)在惰性气体保护下，以含有磷酰胆碱基团的乙烯基单体和含有五氟苯基的乙烯基单体为原料，在引发剂的作用下进行自由基聚合反应，得到含有五氟苯基的磷酰胆碱聚合物；(1) Under the protection of an inert gas, a vinyl monomer containing a phosphorylcholine group and a vinyl monomer containing a pentafluorophenyl group are used as raw materials, and a radical polymerization reaction is carried out under the action of an initiator to obtain a compound containing Phosphorylcholine polymers of pentafluorophenyl;

(2)将所述含有五氟苯基的磷酰胆碱聚合物溶于极性溶剂中，得到聚合物溶液；并加入多巴胺，在惰性气体保护下，于30-55℃进行接枝反应；将所得反应物冷冻干燥，即得所需双仿生聚合物。(2) dissolving the pentafluorophenyl-containing phosphorylcholine polymer in a polar solvent to obtain a polymer solution; and adding dopamine, under the protection of an inert gas, carrying out a grafting reaction at 30-55°C; The obtained reactant is freeze-dried to obtain the desired dual biomimetic polymer.

所述步骤(1)中，所述含有磷酰胆碱基团的乙烯基单体与所述含有五氟苯基的乙烯基单体的摩尔比为3：7-8：2。In the step (1), the molar ratio of the vinyl monomer containing a phosphorylcholine group to the vinyl monomer containing a pentafluorophenyl group is 3:7-8:2.

所述步骤(1)中：In the step (1):

所述含有磷酰胆碱基团的乙烯基单体包括甲基丙烯酰氧乙基磷酰胆碱；The vinyl monomer containing phosphorylcholine group includes methacryloxyethyl phosphorylcholine;

所述含有五氟苯基的乙烯基单体包括甲基丙烯酸五氟苯酯。The pentafluorophenyl-containing vinyl monomer includes pentafluorophenyl methacrylate.

所述步骤(1)中，控制所述自由基聚合反应的温度为60-80℃。In the step (1), the temperature of the radical polymerization reaction is controlled to be 60-80°C.

所述步骤(1)中，还包括将反应原料加入有机溶剂中进行自由基聚合反应的步骤，所述有机溶剂为四氢呋喃，并添加有乙醇或甲醇的混合溶液。所述有机溶剂的用量以实现所述反应原料的充分溶解为宜。In the step (1), it also includes the step of adding the reaction raw materials into an organic solvent to carry out a radical polymerization reaction, the organic solvent is tetrahydrofuran, and a mixed solution of ethanol or methanol is added. The amount of the organic solvent is appropriate to achieve full dissolution of the reaction raw materials.

所述步骤(2)中，所述多巴胺的摩尔量为所述含有五氟苯基的磷酰胆碱聚合物中五氟苯基摩尔量的10-120％。In the step (2), the molar amount of dopamine is 10-120% of the molar amount of pentafluorophenyl in the pentafluorophenyl-containing phosphorylcholine polymer.

所述步骤(2)中，所述极性溶剂包括水、甲醇或乙醇。所述极性溶剂的用量及使得所述反应原料充分溶剂为宜。In the step (2), the polar solvent includes water, methanol or ethanol. The amount of the polar solvent and the sufficient solvent of the reaction raw materials are suitable.

所述步骤(2)中，控制所述接枝反应的温度为30-55℃。In the step (2), the temperature of the grafting reaction is controlled to be 30-55°C.

所述步骤(1)和/或步骤(2)中，还包括将所得反应物进行透析的步骤。In the step (1) and/or step (2), the step of dialyzing the obtained reactant is also included.

本发明还公开了由所述的方法制备得到的双仿生聚合物。The invention also discloses the double biomimetic polymer prepared by the method.

本发明还公开了所述的双仿生聚合物用于进行材料表面生物相容性改性处理的用途。The invention also discloses the use of the dual biomimetic polymer for the modification of the biocompatibility of the material surface.

本发明所述双仿生聚合物以含有磷酰胆碱基团的乙烯基单体和含有五氟苯基的乙烯基单体为原料进行自由基聚合反应，得到含有五氟苯基的磷酰胆碱聚合物，并将仿蚌类粘附蛋白成分多巴胺与含有五氟苯基的磷酰胆碱聚合物相结合，邻二酚羟基具有仿蚌类粘附蛋白多巴胺的粘附作用，赋予水溶性聚合物在材料表面优异的粘附性能，磷酰胆碱具有仿细胞外层膜结构的抗污性能，获得了良好的抗生物污染的性能，该聚合物具有双重仿生的特征，在材料表面修饰领域具有广阔的应用前景。The dual biomimetic polymer of the present invention uses a vinyl monomer containing a phosphorylcholine group and a vinyl monomer containing a pentafluorophenyl group as raw materials to carry out a free radical polymerization reaction to obtain a phosphorylcholine containing a pentafluorophenyl group Alkaline polymer, combining dopamine, a component of mussel-like adhesion proteins, with phosphorylcholine polymers containing pentafluorophenyl, and the ortho-diphenol hydroxyl group has the adhesion of mussel-like adhesion protein dopamine, giving water solubility The polymer has excellent adhesion properties on the surface of the material. Phosphorylcholine has anti-fouling properties that mimic the structure of the outer membrane of cells, and obtains good anti-biological fouling properties. The polymer has dual biomimetic characteristics and is modified on the surface of the material. The field has broad application prospects.

本发明所述双仿生聚合物中的多巴胺基团除具有π-π堆积等多种非共价键作用之外，还易氧化聚合形成粘附性聚多巴胺(PDA)，可与包括金属、玻璃和塑料在内的多种基材产生耐水的强粘附作用。此外，多巴胺涂层可通过迈克尔加成或席夫碱反应接枝具有生物功能的分子。这种仿蚌类粘附的表面修饰方法可以弥补目前物理涂覆必须经过复杂的化学处理才能获得稳定涂层的局限性，简化材料表面改性的条件及过程。而且，本发明所述制备方法利用五氟苯基与氨基反应将多巴胺接枝到含有五氟苯基的磷酰胆碱聚合物上，可以略去酚羟基的保护过程，解决了由于多巴胺单体中的酚羟基是自由基聚合的阻聚剂，导致该类单体聚合时必须进行酚羟基保护的过程，解决了现有多巴胺类聚合物合成的难点问题；而且，本发明方法制备的双仿生聚合物中，不仅所述多巴胺的接枝率可控，且接枝率高达65％，相对于现有技术中一般方法仅为4％的多巴胺的接枝率，本发明制得双仿生聚合物的涂层粘附力进一步增强，并不易脱落。The dopamine group in the dual biomimetic polymer of the present invention not only has various non-covalent bonds such as π-π stacking, but also is easily oxidized and polymerized to form adhesive polydopamine (PDA). Produces strong, water-resistant adhesion to a variety of substrates, including plastics. Furthermore, dopamine coatings can be grafted with biologically functional molecules via Michael addition or Schiff base reactions. This mussel-like adhesion surface modification method can make up for the limitation that the current physical coating must undergo complex chemical treatment to obtain a stable coating, and simplify the conditions and process of material surface modification. Moreover, the preparation method of the present invention utilizes the reaction between pentafluorophenyl and amino groups to graft dopamine onto the phosphorylcholine polymer containing pentafluorophenyl, which can omit the protection process of the phenolic hydroxyl group, and solves the problem that the dopamine monomer The phenolic hydroxyl group is the polymerization inhibitor of free radical polymerization, which leads to the process of protecting the phenolic hydroxyl group during the polymerization of this type of monomer, which solves the difficult problem of the existing dopamine polymer synthesis; In the polymer, not only the grafting rate of the dopamine is controllable, but also the grafting rate is as high as 65%. Compared with the general method in the prior art, the grafting rate of dopamine is only 4%. The double biomimetic polymer is prepared by the present invention. The adhesion of the coating is further enhanced, and it is not easy to fall off.

本发明所述双仿生衍生物，用含有细胞外层膜成分磷酰胆碱和蚌类粘附蛋白成分多巴胺的双仿生聚合物进行改性材料表面，聚合物中的多巴胺侧基从水溶液中可粘附到包括聚四氟乙烯在内的多种材料表面，磷酰胆碱侧基则在涂层表面自动形成仿细胞外层膜结构，显著提高了基底材料的生物相容性。采用本发明方法制备的双仿生聚合物在体内植入材料，组织工程，药物缓释及生物传感器等领域具有广阔的应用前景。The dual biomimetic derivatives of the present invention use dual biomimetic polymers containing phosphorylcholine, a component of the outer cell membrane, and dopamine, a mussel adhesion protein component, to modify the surface of the material, and the dopamine side groups in the polymer can be obtained from an aqueous solution. Adhering to the surface of various materials including polytetrafluoroethylene, the side groups of phosphorylcholine automatically form a cell-like outer membrane structure on the coating surface, which significantly improves the biocompatibility of the substrate material. The dual biomimetic polymer prepared by the method of the present invention has broad application prospects in the fields of in vivo implantation materials, tissue engineering, sustained drug release and biosensors.

本发明所述双仿生衍生物的制备方法，材料来源广泛，制备方法简单，条件温和，且接枝率可控，并解决了现有技术中产物提纯困难的问题，为获得双仿生聚合物提供了一种新的途径。The preparation method of the dual biomimetic derivatives of the present invention has the advantages of wide material sources, simple preparation method, mild conditions, and controllable grafting rate, and solves the problem of difficulty in product purification in the prior art, and provides a method for obtaining dual biomimetic polymers. a new way.

附图说明Description of drawings

为了使本发明的内容更容易被清楚的理解，下面根据本发明的具体实施例并结合附图，对本发明作进一步详细的说明，其中，In order to make the content of the present invention easier to understand clearly, the present invention will be described in further detail below according to specific embodiments of the present invention and in conjunction with the accompanying drawings, wherein,

图1为本发明壳聚糖膜和改性壳聚糖膜的动态接触角，各组中左侧柱状为前进角数值，右侧柱状为后退角数值；Fig. 1 is the dynamic contact angle of the chitosan film of the present invention and the modified chitosan film, in each group, the left column is the advancing angle value, and the right column is the receding angle value;

图2为本发明壳聚糖膜和改性壳聚糖膜表面精细能谱图。Fig. 2 is the surface fine energy spectrogram of the chitosan film of the present invention and the modified chitosan film.

具体实施方式Detailed ways

本发明下述实施例中所述双仿生聚合物的制备基于五氟苯基的接枝反应进行，具体合成路线如下：The preparation of the double biomimetic polymer described in the following examples of the present invention is based on the grafting reaction of pentafluorophenyl, and the specific synthetic route is as follows:

实施例1Example 1

本实施例所述双仿生聚合物的制备方法包括如下步骤：The preparation method of the dual biomimetic polymer described in this embodiment comprises the following steps:

(1)准确称取8mmol2-甲基丙烯酰氧基乙基磷酰胆碱和2mmol甲基丙烯酸五氟苯酯，用乙醇和四氢呋喃混合溶剂(体积比4：1)溶解混合均匀，将0.1mmol偶氮二异丁腈用四氢呋喃溶解得到引发剂溶液，在N₂保护，70℃搅拌条件下，向三颈瓶中加入单体的混合溶液，预热30min后，加入所述引发剂溶液继续反应24h，反应结束后，浓缩反应液，用截留分子量为6000-8000D的透析袋进行透析；最后，将透析后的样品在-50℃下冷冻干燥，即可得到含有五氟苯基的磷酰胆碱聚合物；核磁测试结果表明，该聚合物中五氟苯基的摩尔含量约为17％；(1) Accurately weigh 8mmol of 2-methacryloyloxyethylphosphorylcholine and 2mmol of pentafluorophenyl methacrylate, dissolve and mix homogeneously with ethanol and tetrahydrofuran mixed solvent (volume ratio 4:1). Azobisisobutyronitrile was dissolved in tetrahydrofuran to obtain an initiator solution. Under the protection of N₂ and stirring at 70 °C, the mixed solution of monomers was added to the three-necked flask. After preheating for 30min, the initiator solution was added to continue the reaction. 24h, after the reaction is completed, the reaction solution is concentrated and dialyzed with a dialysis bag with a molecular weight cut-off of 6000-8000D; finally, the dialyzed sample is freeze-dried at -50°C to obtain a phosphoryl bile containing pentafluorophenyl Alkali polymer; NMR test results show that the molar content of pentafluorophenyl in the polymer is about 17%;

用400MHz核磁共振仪以D₂O为溶剂测试聚合物的氢核磁。在5-7ppm处未见出峰，表明所得共聚物中没有残余单体，并成功合成了该聚合物，以3.28ppm处为-N⁺(CH₃)₃特征峰，0.9-2.2ppm处为主链上亚甲基和侧链甲基的峰计算聚合物组成，可知该聚合物组成与投料比基本一致；The hydrogen NMR of the polymer was tested with a 400MHz nuclear magnetic resonance apparatus with D₂ O as the solvent. No peak was seen at 5-7ppm, indicating that there was no residual monomer in the obtained copolymer, and the polymer was successfully synthesized, with a characteristic peak of -N⁺ (CH₃ )₃ at 3.28ppm and a characteristic peak at 0.9-2.2ppm The peaks of methylene and side chain methyl groups on the main chain calculate the polymer composition, and it can be known that the polymer composition is basically consistent with the feeding ratio;

(2)取0.5g上述步骤(1)中制得的所述含有五氟苯基的磷酰胆碱聚合物溶于20mL甲醇中得到聚合物溶液，并在N₂保护，于30℃搅拌条件下，向三颈瓶中加入所述聚合物溶液，预热30min，再加入0.15g多巴胺，进行保温搅拌反应12h；反应结束后，浓缩反应液，用截留分子量为6000-8000D的透析袋在pH值为3-4的盐酸水溶液中透析浓缩后的反应液，将透析后的样品在-50℃下冷冻干燥，即可得到双仿生聚合物。经检测及计算，本实施例所得聚合物中多巴胺的接枝率为11％。(2) Dissolve 0.5 g of the pentafluorophenyl-containing phosphorylcholine polymer prepared in the above step (1) in 20 mL of methanol to obtain a polymer solution, protect under N₂ and stir at 30 °C Then, the polymer solution was added to the three-necked flask, preheated for 30 min, and then 0.15 g of dopamine was added, and the reaction was carried out with heat preservation and stirring for 12 h; The concentrated reaction solution is dialyzed in an aqueous hydrochloric acid solution with a value of 3-4, and the dialyzed sample is freeze-dried at -50° C. to obtain a double biomimetic polymer. After detection and calculation, the graft ratio of dopamine in the polymer obtained in this example was 11%.

实施例2Example 2

(1)称取7mmol2-甲基丙烯酰氧基乙基磷酰胆碱和3mmol甲基丙烯酸五氟苯酯，用甲醇和四氢呋喃混合溶剂溶解混合均匀(体积比4：1)，将0.1mmol偶氮二异丁腈用四氢呋喃溶解得到引发剂溶液，在N₂保护，60℃搅拌条件下，向三颈瓶中加入单体的混合溶液，预热30min后，加入引发剂溶液继续反应24h，反应结束后，浓缩反应液，用截留分子量为6000-8000D的透析袋透析；最后，将透析后的样品在-50℃下冷冻干燥，即可得到含有五氟苯基的磷酰胆碱聚合物；核磁测试结果表明，该聚合物中五氟苯基的摩尔含量约为26％；(1) Weigh 7mmol of 2-methacryloyloxyethylphosphorylcholine and 3mmol of pentafluorophenyl methacrylate, dissolve and mix them in a mixed solvent of methanol and tetrahydrofuran (volume ratio 4:1), add 0.1mmol of The azobisisobutyronitrile was dissolved in tetrahydrofuran to obtain the initiator solution. Under the condition of_N2 protection and stirring at 60 °C, the mixed solution of monomers was added to the three-necked flask. After preheating for 30min, the initiator solution was added to continue the reaction for 24h. After the end, the reaction solution is concentrated and dialyzed with a dialysis bag with a molecular weight cut-off of 6000-8000D; finally, the dialyzed sample is freeze-dried at -50°C to obtain a phosphorylcholine polymer containing pentafluorophenyl; NMR test results show that the molar content of pentafluorophenyl in the polymer is about 26%;

(2)取0.5g上述步骤(1)中制得的所述含有五氟苯基的磷酰胆碱聚合物溶于20mL乙醇中得到聚合物溶液，在N₂保护，于35℃搅拌条件下，向三颈瓶中加入所述聚合物溶液，预热30min，再加入0.2g多巴胺，保温搅拌反应14h；反应结束后，浓缩反应液，用截留分子量为6000-8000D的透析袋在pH值为3-4的盐酸水溶液中透析浓缩后的反应液，将透析后的样品在-50℃下冷冻干燥，即可得到双仿生聚合物。经检测及计算，本实施例所得聚合物中多巴胺的接枝率为19％。(2) Dissolve 0.5 g of the pentafluorophenyl-containing phosphorylcholine polymer prepared in the above step (1) in 20 mL of ethanol to obtain a polymer solution, under the protection of_N2 , under stirring at 35 °C , add the polymer solution to the three-necked flask, preheat for 30min, then add 0.2g dopamine, keep stirring for 14h; after the reaction, concentrate the reaction solution, use a dialysis bag with a molecular weight cut-off of 6000-8000D at pH The concentrated reaction solution was dialyzed in the aqueous hydrochloric acid solution of 3-4, and the dialyzed sample was freeze-dried at -50° C. to obtain the double biomimetic polymer. After detection and calculation, the graft ratio of dopamine in the polymer obtained in this example was 19%.

实施例3Example 3

(1)准确称取6mmol 2-甲基丙烯酰氧基乙基磷酰胆碱和4mmol甲基丙烯酸五氟苯酯，用乙醇和四氢呋喃混合溶剂溶解混合均匀(体积比4：1)，将0.1mmol偶氮二异丁腈用四氢呋喃溶解得到引发剂溶液，在N₂保护，80℃搅拌条件下，向三颈瓶中加入单体的混合溶液，预热30min后，加入引发剂溶液继续反应24h，反应结束后，浓缩反应液，用截留分子量为6000-8000D的透析袋透析；最后，将透析后的样品在-50℃下冷冻干燥，即可得到含有五氟苯基的磷酰胆碱聚合物；核磁测试结果表明，该聚合物中五氟苯基的摩尔含量约为38％；(1) Accurately weigh 6mmol of 2-methacryloyloxyethylphosphorylcholine and 4mmol of pentafluorophenyl methacrylate, dissolve and mix them in a mixed solvent of ethanol and tetrahydrofuran (volume ratio 4:1), mix 0.1 mmol azobisisobutyronitrile was dissolved in tetrahydrofuran to obtain the initiator solution. Under the protection of N₂ and stirring at 80 °C, the mixed solution of monomers was added to the three-necked flask. After preheating for 30min, the initiator solution was added to continue the reaction for 24h. , after the reaction, the reaction solution was concentrated and dialyzed with a dialysis bag with a molecular weight cut-off of 6000-8000D; finally, the dialyzed sample was freeze-dried at -50 °C to obtain a phosphorylcholine polymer containing pentafluorophenyl. NMR test results show that the molar content of pentafluorophenyl in the polymer is about 38%;

(2)取0.5g上述步骤(1)中制得的所述含有五氟苯基的磷酰胆碱聚合物溶于20mL蒸馏水中得到聚合物溶液，在N₂保护，40℃搅拌条件下，向三颈瓶中加入所述聚合物溶液，预热30min，再加入0.4g多巴胺，保温搅拌反应16h；反应结束后，浓缩反应液，用截留分子量为6000-8000D的透析袋在pH值为3-4的盐酸水溶液中透析浓缩后的反应液，将透析后的样品在-50℃下冷冻干燥，即可得到双仿生聚合物。经检测及计算，本实施例所得聚合物中多巴胺的接枝率为31％。(2) Dissolve 0.5 g of the pentafluorophenyl-containing phosphorylcholine polymer prepared in the above step (1) in 20 mL of distilled water to obtain a polymer solution, under the protection of_N2 , under stirring conditions of 40 °C, The polymer solution was added to the three-necked flask, preheated for 30 min, and then 0.4 g of dopamine was added, and the reaction was kept under stirring for 16 h; after the reaction was completed, the reaction solution was concentrated, and a dialysis bag with a molecular weight cut-off of 6000-8000 D was used at a pH of 3. The concentrated reaction solution was dialyzed in an aqueous hydrochloric acid solution of -4, and the dialyzed sample was freeze-dried at -50°C to obtain a double biomimetic polymer. After detection and calculation, the graft ratio of dopamine in the polymer obtained in this example was 31%.

实施例4Example 4

(1)称取5mmol 2-甲基丙烯酰氧基乙基磷酰胆碱和5mmol甲基丙烯酸五氟苯酯，用甲醇和四氢呋喃混合溶剂溶解混合均匀(体积比4：1)，将0.1mmol偶氮二异丁腈用四氢呋喃溶解得到引发剂溶液，在N₂保护，65℃搅拌条件下，向三颈瓶中加入单体的混合溶液，预热30min后，加入引发剂溶液继续反应24h，反应结束后，浓缩反应液，用截留分子量为6000-8000D的透析袋透析；最后，将透析后的样品在-50℃下冷冻干燥，即可得到含有五氟苯基的磷酰胆碱聚合物；核磁测试结果表明，该聚合物中五氟苯基的摩尔含量约为47％；(1) Weigh 5mmol of 2-methacryloyloxyethylphosphorylcholine and 5mmol of pentafluorophenyl methacrylate, dissolve and mix them with methanol and tetrahydrofuran mixed solvent (volume ratio 4:1), add 0.1mmol Azobisisobutyronitrile was dissolved in tetrahydrofuran to obtain the initiator solution. Under the protection of N₂ and stirring at 65 °C, the mixed solution of monomers was added to the three-necked flask. After preheating for 30min, the initiator solution was added to continue the reaction for 24h. After the reaction is completed, the reaction solution is concentrated and dialyzed with a dialysis bag with a molecular weight cut-off of 6000-8000D; finally, the dialyzed sample is freeze-dried at -50°C to obtain a phosphorylcholine polymer containing pentafluorophenyl ; NMR test results show that the molar content of pentafluorophenyl in the polymer is about 47%;

(2)取0.5g步骤(1)中得到的所述含有五氟苯基的磷酰胆碱聚合物溶于20mL甲醇中得到聚合物溶液，在N₂保护，45℃搅拌条件下，向三颈瓶中加入所述聚合物溶液，预热30min，再加入0.2g多巴胺，保温搅拌反应18h。反应结束后，浓缩反应液，用截留分子量为6000-8000D的透析袋在pH值为3-4的盐酸水溶液中透析浓缩后的反应液，将透析后的样品在-50℃下冷冻干燥，即可得到双仿生聚合物。经检测及计算，本实施例所得聚合物中多巴胺的接枝率为40％。(2) Dissolve 0.5 g of the pentafluorophenyl-containing phosphorylcholine polymer obtained in step (1) in 20 mL of methanol to obtain a polymer solution, under N₂ protection, under stirring conditions at 45 °C, to three The polymer solution was added to the neck flask, preheated for 30 minutes, then 0.2 g of dopamine was added, and the reaction was kept under stirring for 18 hours. After the reaction is completed, the reaction solution is concentrated, and the concentrated reaction solution is dialyzed in an aqueous hydrochloric acid solution with a pH value of 3-4 using a dialysis bag with a molecular weight cut-off of 6000-8000D, and the dialyzed sample is freeze-dried at -50 °C, that is, Dual biomimetic polymers are available. After detection and calculation, the graft ratio of dopamine in the polymer obtained in this example was 40%.

实施例5Example 5

(1)称取4mmol 2-甲基丙烯酰氧基乙基磷酰胆碱和6mmol甲基丙烯酸五氟苯酯，用甲醇和四氢呋喃混合溶剂溶解混合均匀(体积比4：1)，将0.1mmol偶氮二异丁腈用四氢呋喃溶解得到引发剂溶液，在N₂保护，75℃搅拌条件下，向三颈瓶中加入单体的混合溶液，预热30min后，加入引发剂溶液继续反应24h，反应结束后，浓缩反应液，用截留分子量为6000-8000D的透析袋透析；最后，将透析后的样品在-50℃下冷冻干燥，即可得到含有五氟苯基的磷酰胆碱聚合物；核磁测试结果表明，该聚合物中五氟苯基的摩尔含量约为57％；(1) Weigh 4mmol of 2-methacryloyloxyethylphosphorylcholine and 6mmol of pentafluorophenyl methacrylate, dissolve and mix them with methanol and tetrahydrofuran mixed solvent (volume ratio 4:1), mix 0.1mmol Azobisisobutyronitrile was dissolved in tetrahydrofuran to obtain an initiator solution. Under the protection of N₂ and stirring at 75 °C, the mixed solution of monomers was added to the three-necked flask. After preheating for 30min, the initiator solution was added to continue the reaction for 24h. After the reaction is completed, the reaction solution is concentrated and dialyzed with a dialysis bag with a molecular weight cut-off of 6000-8000D; finally, the dialyzed sample is freeze-dried at -50°C to obtain a phosphorylcholine polymer containing pentafluorophenyl ; NMR test results show that the molar content of pentafluorophenyl in the polymer is about 57%;

(2)取0.5g步骤(1)中所述含有五氟苯基的磷酰胆碱聚合物溶于20mL乙醇中得到聚合物溶液，在N₂保护，50℃搅拌条件下，向三颈瓶中加入所述聚合物溶液，预热30min，再加入0.5g多巴胺，保温搅拌反应20h。反应结束后，浓缩反应液，用截留分子量为6000-8000D的透析袋在pH值为3-4的盐酸水溶液中透析浓缩后的反应液，将透析后的样品在-50℃下冷冻干燥，即可得到双仿生聚合物。经检测及计算，本实施例所得聚合物中多巴胺的接枝率为49％。(2) Dissolve 0.5 g of the phosphorylcholine polymer containing pentafluorophenyl described in step (1) in 20 mL of ethanol to obtain a polymer solution, under N₂ protection, under stirring conditions of 50 ° C, to a three-necked flask The polymer solution was added to the solution, preheated for 30 min, and then 0.5 g of dopamine was added, and the reaction was kept under stirring for 20 h. After the reaction is completed, the reaction solution is concentrated, and the concentrated reaction solution is dialyzed in an aqueous hydrochloric acid solution with a pH value of 3-4 using a dialysis bag with a molecular weight cut-off of 6000-8000D, and the dialyzed sample is freeze-dried at -50 °C, that is, Dual biomimetic polymers are available. After detection and calculation, the graft ratio of dopamine in the polymer obtained in this example was 49%.

实施例6Example 6

(1)称取3mmol 2-甲基丙烯酰氧基乙基磷酰胆碱和7mmol甲基丙烯酸五氟苯酯，用甲醇和四氢呋喃混合溶剂溶解混合均匀(体积比4：1)，将0.1mmol偶氮二异丁腈用四氢呋喃溶解得到引发剂溶液，在N₂保护，70℃搅拌条件下，向三颈瓶中加入单体的混合溶液，预热30min后，加入引发剂溶液继续反应24h，反应结束后，浓缩反应液，用截留分子量为6000-8000D的透析袋透析；最后，将透析后的样品在-50℃下冷冻干燥，即可得到含有五氟苯基的磷酰胆碱聚合物；核磁测试结果表明，该聚合物中五氟苯基的摩尔含量约为71％；(1) Weigh 3mmol of 2-methacryloyloxyethylphosphorylcholine and 7mmol of pentafluorophenyl methacrylate, dissolve and mix them with methanol and tetrahydrofuran mixed solvent (volume ratio 4:1), add 0.1mmol Azobisisobutyronitrile was dissolved in tetrahydrofuran to obtain the initiator solution. Under the protection of N₂ and stirring at 70 °C, the mixed solution of monomers was added to the three-necked flask. After preheating for 30min, the initiator solution was added to continue the reaction for 24h. After the reaction, the reaction solution was concentrated and dialyzed with a dialysis bag with a molecular weight cut-off of 6000-8000D; finally, the dialyzed sample was freeze-dried at -50°C to obtain a pentafluorophenyl-containing phosphorylcholine polymer ; NMR test results show that the molar content of pentafluorophenyl in the polymer is about 71%;

(2)取0.5g步骤(1)中所述含有五氟苯基的磷酰胆碱聚合物溶于20mL蒸馏水中得到聚合物溶液，在N₂保护，55℃搅拌条件下，向三颈瓶中加入所述聚合物溶液，预热30min，再加入0.7g多巴胺，保温搅拌反应24h。反应结束后，浓缩反应液，用截留分子量为6000-8000D的透析袋在pH值为3-4的盐酸水溶液中透析浓缩后的反应液，将透析后的样品在-50℃下冷冻干燥，即可得到双仿生聚合物。经检测及计算，本实施例所得聚合物中多巴胺的接枝率为65％。(2) Dissolve 0.5 g of the phosphorylcholine polymer containing pentafluorophenyl described in step (1) in 20 mL of distilled water to obtain a polymer solution, under N₂ protection, under stirring conditions of 55 ° C, to a three-necked flask The polymer solution was added to the solution, preheated for 30 min, and then 0.7 g of dopamine was added, and the reaction was maintained and stirred for 24 h. After the reaction is completed, the reaction solution is concentrated, and the concentrated reaction solution is dialyzed in an aqueous hydrochloric acid solution with a pH value of 3-4 using a dialysis bag with a molecular weight cut-off of 6000-8000D, and the dialyzed sample is freeze-dried at -50 °C, that is, Dual biomimetic polymers are available. After detection and calculation, the graft ratio of dopamine in the polymer obtained in this example was 65%.

本发明所述双仿生聚合物可用于改性材料表面的生物相容性，具体应用为通过将制得的双仿生聚合物溶于极性溶剂中，并将其滴凃在待改性材料膜表面，待晾干后置于pH＝8.0-9.0水溶液中于30-80℃处理，依靠磷酰胆碱聚合物中五氟苯基与壳聚糖膜表面氨基亲核取代反应的锚定作用以及多巴胺的粘附将磷酰胆碱基团固定在壳聚糖膜表面，即可制备具有仿细胞外层膜结构的粘附仿生涂层。The dual biomimetic polymer of the present invention can be used to modify the biocompatibility of the surface of the material, and the specific application is by dissolving the prepared dual biomimetic polymer in a polar solvent and drop-coating it on the film of the material to be modified. The surface, after drying, is placed in an aqueous solution of pH=8.0-9.0 and treated at 30-80 °C, relying on the anchoring effect of the pentafluorophenyl in the phosphorylcholine polymer and the nucleophilic substitution reaction of the amino group on the surface of the chitosan film and the The adhesion of dopamine fixes the phosphorylcholine group on the surface of the chitosan film, and then the adhesion biomimetic coating with the structure of the outer layer of the cell can be prepared.

如图1所示，本实施例经涂层处理的壳聚糖膜与未经涂层处理的壳聚糖膜相比，经涂层处理的壳聚糖的前进角和后退角均有所降低，这是因为亲水性好的磷酰胆碱聚合物通过五氟苯基与壳聚糖表面氨基的反应锚定以及多巴胺的粘附将磷酰胆碱基团粘附固定在壳聚糖膜的表面，获得具有仿细胞外层膜结构的表面，使得其亲水性显著提高，前进角和后退角明显降低。As shown in Figure 1, the advancing angle and the receding angle of the coated chitosan film are reduced compared with the uncoated chitosan film in this example. , this is because the phosphorylcholine polymer with good hydrophilicity is anchored by the reaction of pentafluorophenyl group with amino groups on the surface of chitosan and the adhesion of dopamine to fix the phosphorylcholine group on the chitosan film , the surface with the structure of the outer membrane of the cell is obtained, so that its hydrophilicity is significantly improved, and the advancing angle and the receding angle are significantly reduced.

如图2所示，本实施例经涂层处理的壳聚糖膜与未经涂层处理的壳聚糖膜相比，经改性处理的壳聚糖膜表面有磷酰胆碱基团上N和P特征吸收峰，这说明亲水性好的磷酰胆碱基团被固定在壳聚糖膜表面。通过五氟苯基与壳聚糖表面氨基的反应锚定以及多巴胺的粘附将磷酰胆碱基团粘附固定在壳聚糖膜的表面，使得其亲水性显著提高，出现磷酰胆碱基团上N和P特征吸收峰。可见，本发明所述方法能够对壳聚糖材料表面的生物相容性进行有效的改善。As shown in Figure 2, compared with the uncoated chitosan film, the modified chitosan film has phosphorylcholine groups on the surface of the coated chitosan film. The characteristic absorption peaks of N and P indicate that the hydrophilic phosphorylcholine groups are immobilized on the surface of the chitosan film. The phosphorylcholine group is adhered and fixed on the surface of the chitosan film through the reaction anchoring of pentafluorophenyl and the amino group on the surface of chitosan and the adhesion of dopamine, which makes the hydrophilicity significantly improved, and the phosphorylcholine appears. The characteristic absorption peaks of N and P on the base group. It can be seen that the method of the present invention can effectively improve the biocompatibility of the surface of the chitosan material.

显然，上述实施例仅仅是为清楚地说明所作的举例，而并非对实施方式的限定。对于所属领域的普通技术人员来说，在上述说明的基础上还可以做出其它不同形式的变化或变动。这里无需也无法对所有的实施方式予以穷举。而由此所引伸出的显而易见的变化或变动仍处于本发明创造的保护范围之中。Obviously, the above-mentioned embodiments are only examples for clear description, and are not intended to limit the implementation manner. For those of ordinary skill in the art, changes or modifications in other different forms can also be made on the basis of the above description. There is no need and cannot be exhaustive of all implementations here. And the obvious changes or changes derived from this are still within the protection scope of the present invention.

Claims

1. The preparation method of the double bionic polymer is characterized by comprising the following steps:

(1) under the protection of inert gas, taking a vinyl monomer containing a phosphorylcholine group and a vinyl monomer containing a pentafluorophenyl group as raw materials, and carrying out free radical polymerization reaction under the action of an initiator to obtain a phosphorylcholine polymer containing the pentafluorophenyl group;

(2) dissolving the phosphorylcholine polymer containing pentafluorophenyl into a polar solvent to obtain a polymer solution; under the protection of inert gas, adding dopamine to carry out grafting reaction; freeze-drying the obtained reactant to obtain the required double-bionic polymer;

in the step (1), the molar ratio of the vinyl monomer containing the phosphorylcholine group to the vinyl monomer containing the pentafluorophenyl group is 3:7-8: 2;

in the step (2), the molar amount of the dopamine is 10-120% of the molar amount of pentafluorophenyl group in the phosphorylcholine polymer containing pentafluorophenyl group.

2. The method for preparing a biomimetic polymer according to claim 1, wherein in the step (1): the vinyl monomer containing phosphorylcholine groups comprises methacryloyloxyethyl phosphorylcholine; the pentafluorophenyl containing vinyl monomer includes pentafluorophenyl methacrylate.

3. The method for preparing a biomimetic polymer according to claim 1 or 2, wherein in the step (1), the temperature of the radical polymerization reaction is controlled to be 60-80 ℃.

4. The method for preparing the biomimetic polymer according to claim 1 or 2, wherein the step (1) further comprises a step of adding the reaction raw material into an organic solvent to perform a radical polymerization reaction, wherein the organic solvent is tetrahydrofuran and a mixed solution of ethanol or methanol is added.

5. The method for preparing a biomimetic polymer according to claim 1 or 2, wherein in the step (2), the polar solvent comprises water, methanol or ethanol.

6. The method for preparing a biomimetic polymer according to claim 1 or 2, wherein in the step (2), the temperature of the grafting reaction is controlled to be 30-55 ℃.

7. A bionical polymer produced by the method of any one of claims 1-6.

8. Use of the biomimetic polymer of claim 7 for performing a material surface biocompatibility modification treatment.