CN1305916C

Movatterモバイル変換

Info

Publication number: CN1305916C
Application number: CNB2004100613153A
Authority: CN
Inventors: 卓仁禧; 张建涛; 黄世文
Original assignee: Wuhan University WHU
Current assignee: Wuhan University WHU
Priority date: 2004-12-10
Filing date: 2004-12-10
Publication date: 2007-03-21
Anticipated expiration: 2024-12-10
Also published as: CN1648141A

Abstract

含硅氧烷侧链的温度敏感线性聚合物：以N-异丙基丙烯酰胺和(γ-甲基丙烯酰基)丙基三甲氧基硅烷作为单体，溶于1，4-二氧六环，以过氧化苯甲酰为引发剂在65-90℃引发聚合反应2-24小时，得到温度敏感线性聚合物。将上述聚合物溶于水调pH值3-12，水解交联反应12-72小时，得到温度敏感水凝胶。本发明聚合物合成简单；凝胶合成条件温和(pH值接近中性)，合成凝胶产生的小分子醇的数量少，不会破坏生物分子的活性；凝胶的溶胀性能可以通过控制制备条件调控；所得凝胶具有刺激响应性能。本发明所得水凝胶可以很好地负载生物活性分子，且可以保持生物分子的活性，可以用于生物活性分子的负载、固定化载体或药物控制释放载体。

Temperature-sensitive linear polymer containing siloxane side chains: N-isopropylacrylamide and (γ-methacryloyl)propyltrimethoxysilane as monomers, dissolved in 1,4-dioxane , using benzoyl peroxide as an initiator to initiate a polymerization reaction at 65-90° C. for 2-24 hours to obtain a temperature-sensitive linear polymer. The above polymer is dissolved in water to adjust the pH value to 3-12, and the hydrolysis and cross-linking reaction is carried out for 12-72 hours to obtain a temperature-sensitive hydrogel. The polymer of the present invention is easy to synthesize; the gel synthesis condition is mild (pH value is close to neutral), and the amount of small molecular alcohol produced by the synthetic gel is small, which will not destroy the activity of biomolecules; the swelling performance of the gel can be controlled by the preparation conditions regulation; the resulting gel has stimuli-responsive properties. The hydrogel obtained in the present invention can well load bioactive molecules and maintain the activity of biomolecules, and can be used for bioactive molecule loading, immobilized carrier or drug controlled release carrier.

Description

Translated fromChinese

含硅氧烷侧链的温度敏感线性聚合物及其水凝胶和制法Temperature-sensitive linear polymers containing siloxane side chains, their hydrogels and their preparation methods

技术领域technical field

本发明涉及一种含硅氧烷侧链的温度敏感线性聚合物及其水凝胶和制法。The invention relates to a temperature-sensitive linear polymer containing siloxane side chain, its hydrogel and its preparation method.

背景技术Background technique

水凝胶是一种能大量吸水溶胀的三维聚合物网络结构，一般由亲水性的均聚物或共聚物交联而成，由于物理或化学交联点的存在，所以水凝胶不溶于水。在过去的三十多年中，关于环境敏感和生理敏感水凝胶的研究大量涌现，此类凝胶引起了研究者的广泛关注。环境敏感水凝胶可以对外界环境的微小变化如温度、pH等产生应答，凝胶的性质发生巨大转变，出现相转变，宏观上表现为体积收缩或溶胀。众多学者已将这种独特的性质用于设计各种各样的生物医用装置，如药物释放系统、传感器、人工肌肉、酶的固定化载体、基因传输系统等。Hydrogel is a three-dimensional polymer network structure that can absorb a large amount of water and swell. It is generally formed by cross-linking hydrophilic homopolymers or copolymers. Due to the existence of physical or chemical cross-linking points, hydrogels are insoluble in water. In the past three decades, a large number of studies on environmentally sensitive and physiologically sensitive hydrogels have emerged, and such gels have attracted extensive attention from researchers. Environmentally sensitive hydrogels can respond to small changes in the external environment such as temperature and pH, and the properties of the gel undergo a huge change, such as phase transition, which is manifested as volume shrinkage or swelling macroscopically. Many scholars have used this unique property to design a variety of biomedical devices, such as drug delivery systems, sensors, artificial muscles, enzyme immobilization carriers, gene delivery systems, etc.

在众多的环境敏感水凝胶中，温度敏感水凝胶是最令人关注的一类。聚(N-异丙基丙烯酰胺)(PNIPA)水凝胶是一种典型的温度敏感水凝胶，其在32℃～34℃之间有一个相转变温度，称为较低临界溶解温度(Lower Critical Solution Temperature，LCST)。当外界温度低于LCST时，PNIPA水凝胶吸水溶胀，此时水凝胶为无色透明；而当外界温度高于LCST时，水凝胶剧烈收缩失水，发生相分离。Among the numerous environment-sensitive hydrogels, temperature-sensitive hydrogels are the most interesting one. Poly(N-isopropylacrylamide) (PNIPA) hydrogel is a typical temperature-sensitive hydrogel, which has a phase transition temperature between 32°C and 34°C, called the lower critical solution temperature ( Lower Critical Solution Temperature, LCST). When the external temperature is lower than the LCST, the PNIPA hydrogel absorbs water and swells, and the hydrogel is colorless and transparent; when the external temperature is higher than the LCST, the hydrogel shrinks violently and loses water, and phase separation occurs.

人们利用敏感型智能材料来设计各种各样的药物传输系统，通过调节外界的环境参数来控制药物的释放动力学，而温度敏感水凝胶可能是应用的最为广泛的一类环境敏感材料。由于温度敏感水凝胶可以对外界环境的温度变化产生应答，发生收缩或溶胀，产生“开-关”效应，根据这种特点，研究者设计了不同的释放装置。温度敏感水凝胶用于设计蛋白药物的释放体系时，有以下优点：1.保护药物免受外界恶劣环境，如酶解、低pH的破坏；2.控制人体对药物的反应；3.通过控制外界环境调节凝胶的结构，进一步做到“按需释放”。Peppas等用EGDMA为交联剂，合成了NIPA和甲基丙烯酸(MAAc)的共聚凝胶，用于肝素在血栓位置的刺激释放，研究者也实现了溶栓酶的脉冲释放。Kim等最近报道了P(NIPA-co-BMA-co-AAc)三聚物凝胶释放人降血钙素用于治疗骨质疏松症和血钙过多。Park研究了温度/pH敏感水凝胶对胰岛素的释放，发现药物释放曲线和凝胶孔径有关。People use sensitive smart materials to design various drug delivery systems, and control the release kinetics of drugs by adjusting external environmental parameters, and temperature-sensitive hydrogels may be the most widely used type of environment-sensitive materials. Because the temperature-sensitive hydrogel can respond to the temperature change of the external environment, shrink or swell, and produce an "on-off" effect, according to this characteristic, the researchers designed different release devices. When the temperature-sensitive hydrogel is used to design the release system of protein drugs, it has the following advantages: 1. Protect the drug from the external harsh environment, such as enzymatic hydrolysis and low pH damage; 2. Control the human body's response to the drug; 3. Through Control the external environment to adjust the structure of the gel, and further achieve "on-demand release". Peppas et al. used EGDMA as a cross-linking agent to synthesize a copolymer gel of NIPA and methacrylic acid (MAAc) for the stimulated release of heparin at the thrombus site. The researchers also realized the pulsed release of thrombolytic enzyme. Kim et al recently reported that P(NIPA-co-BMA-co-AAc) terpolymer gel released human calcitonin for the treatment of osteoporosis and hypercalcemia. Park studied the release of insulin from temperature/pH sensitive hydrogels and found that the drug release curve was related to the gel pore size.

水凝胶用于释放活性大分子药物时，同样也存在着一些问题，如负载药物时，若将单体和蛋白药物混合后引发反应，反应过程中产生的高能物质如自由基等会使蛋白变性，并且药物分子很难从凝胶内部完全释放出来；若将合成好的凝胶放入药液中浸泡吸收，由于尺寸排除效应的影响，使得载药率非常低。When the hydrogel is used to release active macromolecular drugs, there are also some problems. For example, when the drug is loaded, if the monomer and the protein drug are mixed to trigger a reaction, the high-energy substances such as free radicals generated during the reaction will cause the protein to Denaturation, and it is difficult for the drug molecules to be completely released from the inside of the gel; if the synthesized gel is immersed in the drug solution for absorption, the drug loading rate is very low due to the size exclusion effect.

发明内容Contents of the invention

本发明所要解决的问题是提供一种含硅氧烷侧链的温度敏感线性聚合物及其水凝胶和制法，该方法条件温和，所得水凝胶可以很好地负载生物活性分子，且可以保持生物分子的活性，可以用于生物活性分子的固定化载体或药物释放载体。The problem to be solved by the present invention is to provide a temperature-sensitive linear polymer containing siloxane side chains and its hydrogel and its preparation method. The method has mild conditions, and the obtained hydrogel can well load biologically active molecules, and It can maintain the activity of biomolecules, and can be used as an immobilized carrier or a drug release carrier for bioactive molecules.

本发明提供的技术方案是：含硅氧烷侧链的温度敏感线性聚合物，由下法制得：按99g∶1ml-1g∶50ml的比例取N-异丙基丙烯酰胺和(γ-甲基丙烯酰基)丙基三甲氧基硅烷作为单体，并按1g∶1-10ml的比例将单体溶于1，4-二氧六环，以过氧化苯甲酰为引发剂在65-90℃引发聚合反应2-24小时，得到分子量为5000-100,000的含硅氧烷侧链的温度敏感线性聚合物。The technical scheme provided by the invention is: a temperature-sensitive linear polymer containing siloxane side chains, which is prepared by the following method: take N-isopropylacrylamide and (γ-methyl Acryloyl) propyltrimethoxysilane is used as a monomer, and the monomer is dissolved in 1,4-dioxane at a ratio of 1g: 1-10ml, and benzoyl peroxide is used as an initiator at 65-90°C The polymerization reaction is initiated for 2-24 hours to obtain a temperature-sensitive linear polymer containing siloxane side chains with a molecular weight of 5000-100,000.

上述单体与引发剂的用量重量比为100∶0.2-5。The weight ratio of the above-mentioned monomer to the initiator is 100:0.2-5.

温度敏感水凝胶，由下法制得：将上述含硅氧烷侧链的温度敏感线性聚合物溶于水制得浓度为5-60wt％的溶液，然后加入磷酸盐/柠檬酸盐的混合溶液至溶液的pH值3-12，水解交联反应12-72小时，得到温度敏感水凝胶。The temperature-sensitive hydrogel is prepared by the following method: dissolving the above-mentioned temperature-sensitive linear polymer containing siloxane side chains in water to obtain a solution with a concentration of 5-60 wt%, and then adding a mixed solution of phosphate/citrate The pH value of the solution is 3-12, and the hydrolysis and cross-linking reaction is carried out for 12-72 hours to obtain a temperature-sensitive hydrogel.

本发明聚合物合成简单；凝胶合成条件温和(pH值接近中性)，合成凝胶产生的小分子醇的数量少，不会破坏生物分子的活性；凝胶的溶胀性能可以通过控制制备条件调控；所得凝胶具有刺激响应性能。本发明所得水凝胶可以很好地负载生物活性分子，且可以保持生物分子的活性，可以用于生物活性分子的负载、固定化载体或药物控制释放载体。The polymer of the present invention is easy to synthesize; the gel synthesis condition is mild (pH value is close to neutral), and the amount of small molecular alcohol produced by the synthetic gel is small, which will not destroy the activity of biomolecules; the swelling performance of the gel can be controlled by the preparation conditions regulation; the resulting gel has stimuli-responsive properties. The hydrogel obtained in the present invention can well load bioactive molecules and maintain the activity of biomolecules, and can be used for bioactive molecule loading, immobilized carrier or drug controlled release carrier.

附图说明：Description of drawings:

图1是本发明所得聚[N-异丙基丙烯酰胺-co-(γ-甲基丙烯酰基)丙基三甲氧基硅烷]的HNMR谱图；Fig. 1 is the HNMR spectrogram of poly[N-isopropylacrylamide-co-(γ-methacryloyl) propyltrimethoxysilane] of the present invention;

图2是本发明中所合成凝胶的溶胀率对温度的关系曲线，说明凝胶的温度敏感性；Fig. 2 is the relational curve of the swelling rate of the gel synthesized in the present invention to temperature, illustrates the temperature sensitivity of gel;

图3是活性蛋白从凝胶释放动力学曲线；Fig. 3 is active protein release kinetics curve from gel;

图4是释放出的小牛血清蛋白的凝胶电泳图。Fig. 4 is a gel electrophoresis image of released bovine serum albumin.

具体实施方式Detailed ways

1.聚[N-异丙基丙烯酰胺-co-(γ-甲基丙烯酰基)丙基三甲氧基硅烷]的合成1. Synthesis of poly[N-isopropylacrylamide-co-(γ-methacryloyl)propyltrimethoxysilane]

原料及投料量：Raw material and feeding quantity:

N-异丙基丙烯酰胺(g)∶(γ-甲基丙烯酰基)丙基三甲氧基硅烷(ml)＝99∶1-1∶50(简称单体)N-isopropylacrylamide (g): (γ-methacryloyl) propyltrimethoxysilane (ml) = 99:1-1:50 (monomer for short)

单体(g)∶过氧化苯甲酰(g)＝100∶0.2-100∶5Monomer (g): benzoyl peroxide (g) = 100:0.2-100:5

单体(g)∶二氧六环(ml)＝1∶1-10Monomer (g): dioxane (ml) = 1: 1-10

取N-异丙基丙烯酰胺和(γ-甲基丙烯酰)丙基三甲氧基硅烷溶于二氧六环，加入过氧化苯甲酰，通氮气15-60min，于65-90℃下引发反应2-24小时。旋转蒸发溶剂，用四氢呋喃溶解，乙醚沉降，干燥；再用四氢呋喃溶解，石油醚沉降，干燥备用。Dissolve N-isopropylacrylamide and (γ-methacryloyl)propyltrimethoxysilane in dioxane, add benzoyl peroxide, blow nitrogen for 15-60min, and trigger at 65-90°C Reaction 2-24 hours. The solvent was evaporated by rotary evaporation, dissolved in tetrahydrofuran, settled in diethyl ether, and dried; then dissolved in tetrahydrofuran, settled in petroleum ether, and dried for later use.

2.温度敏感水凝胶的合成2. Synthesis of temperature-sensitive hydrogels

将聚[N-异丙基丙烯酰胺-co-(γ-甲基丙烯酰基)丙基三甲氧基硅烷]溶于水得到一定浓度(质量百分比5％-60％)的溶液，然后加入磷酸盐/柠檬酸盐的混合溶液，调整溶液的pH值3-12，水解交联反应进行12-72小时。Dissolve poly[N-isopropylacrylamide-co-(γ-methacryloyl)propyltrimethoxysilane] in water to obtain a solution with a certain concentration (5%-60% by mass), and then add phosphate / citrate mixed solution, adjust the pH value of the solution to 3-12, and carry out the hydrolysis and cross-linking reaction for 12-72 hours.

3.以温度敏感水凝胶为载体对生物活性分子的负载和释放3. Loading and release of bioactive molecules using temperature-sensitive hydrogel as a carrier

将聚[N-异丙基丙烯酰胺-co-(γ-甲基丙烯酰基)丙基三甲氧基硅烷]溶于水，加入一定量的生物活性大分子(聚合物∶生物活性大分子＝10∶0.5-5)；然后加入磷酸盐/柠檬酸盐的混合溶液，调整溶液的pH值3-12，水解交联反应进行12-72小时得到含生物活性分子的凝胶。将上述凝胶放入一定量的磷酸盐磷酸盐缓冲溶液，测定释放出来的生物活性分子数量和活性。Poly[N-isopropylacrylamide-co-(γ-methacryloyl)propyltrimethoxysilane] was dissolved in water, and a certain amount of bioactive macromolecule was added (polymer: bioactive macromolecule=10 : 0.5-5); then add the mixed solution of phosphate/citrate, adjust the pH value of the solution to 3-12, carry out the hydrolysis and cross-linking reaction for 12-72 hours to obtain the gel containing bioactive molecules. The above gel is put into a certain amount of phosphate buffered saline solution, and the quantity and activity of released biologically active molecules are measured.

本发明采用两步法制备生物分子载体，先合成的线性聚合物具有水溶性、无毒、可控制分子量等优点；水解交联成凝胶时，在非常温和的条件下(室温，生理pH值)进行，产生的副产物小分子醇浓度极低，因而可以保持活性大分子的活性，且负载率和释放速率可以调控。The present invention adopts two-step method to prepare biomolecule carrier, and the linear polymer synthesized earlier has advantages such as water solubility, nontoxicity, controllable molecular weight; ), the concentration of the by-product small molecule alcohol is extremely low, so the activity of the active macromolecule can be maintained, and the loading rate and release rate can be adjusted.

实施例1：Example 1:

取N-异丙基丙烯酰胺2g，(γ-甲基丙烯酰)丙基三甲氧基硅烷0.3mL溶于15ml二氧六环，加入50mg过氧化苯甲酰，通氮气30min，于80℃下引发反应4小时。旋转蒸发溶剂，用四氢呋喃溶解，乙醚沉降，干燥；再用四氢呋喃溶解，石油醚沉降，干燥得到聚[N-异丙基丙烯酰胺-co-(γ-甲基丙烯酰基)丙基三甲氧基硅烷]。聚合物分子量45000，分散度3.2。核磁图谱见图1：δ3.90-4.05(m，CH(CH₃)₂+OCH₂(CH₂)₂)，3.47(s，Si(OCH₃)₃)，2.1-2.3(d，CHCO)，1.83(d，CH₂CH₂CH₂-)，1.6-1.7(m，CH₂CHCH₂)，1.0-1.2(d，CH(CH₃)₂，C-CH₃).Take 2g of N-isopropylacrylamide, dissolve 0.3mL of (γ-methacryloyl)propyltrimethoxysilane in 15ml of dioxane, add 50mg of benzoyl peroxide, blow nitrogen for 30min, and set the temperature at 80°C The reaction was initiated for 4 hours. Evaporate the solvent by rotary evaporation, dissolve in THF, settle in ether, and dry; dissolve in THF again, settle in petroleum ether, and dry to obtain poly[N-isopropylacrylamide-co-(γ-methacryloyl)propyltrimethoxysilane ]. The molecular weight of the polymer is 45000, and the degree of dispersion is 3.2. See Figure 1 for NMR spectrum: δ3.90-4.05(m, CH(CH₃ )₂ +OCH₂ (CH₂ )₂ ), 3.47(s, Si(OCH₃ )₃ ), 2.1-2.3(d, CHCO) , 1.83 (d, CH₂ CH₂ CH₂ -), 1.6-1.7 (m, CH₂ CHCH₂ ), 1.0-1.2 (d, CH(CH₃ )₂ , C-CH₃ ).

2.温度敏感水凝胶的合成2. Synthesis of temperature-sensitive hydrogels

取聚[N-异丙基丙烯酰胺-co-(γ-甲基丙烯酰基)丙基三甲氧基硅烷]200mg，加入1ml水，于室温下溶解，然后加入0.5ml磷酸盐/柠檬酸缓冲溶液，使反应液的pH值为6，反应于室温下进行24h，取出凝胶在水中浸泡除去杂质。将凝胶在不同温度的水中放置24h，取出称重，测定凝胶的温度敏感性。图2是凝胶溶胀率和温度的关系图，可看出凝胶在低温下溶胀，高温下收缩，其相变温度在29℃。Take poly[N-isopropylacrylamide-co-(γ-methacryloyl)propyltrimethoxysilane] 200mg, add 1ml water, dissolve at room temperature, then add 0.5ml phosphate/citric acid buffer solution , the pH value of the reaction solution was 6, and the reaction was carried out at room temperature for 24 hours, and the gel was taken out and soaked in water to remove impurities. Place the gel in water at different temperatures for 24 hours, take it out and weigh it, and measure the temperature sensitivity of the gel. Figure 2 is a graph showing the relationship between gel swelling rate and temperature. It can be seen that the gel swells at low temperature and shrinks at high temperature, and its phase transition temperature is 29°C.

实施例2：Example 2:

取N-异丙基丙烯酰胺2g，(γ-甲基丙烯酰)丙基三甲氧基硅烷3mL溶于50ml二氧六环，加入100mg过氧化苯甲酰，通氮气60min，于65℃下引发反应24小时。旋转蒸发溶剂，用四氢呋喃溶解，乙醚沉降，干燥；再用四氢呋喃溶解，石油醚沉降，干燥备用，聚合物分子量41000，分散度3.0；核磁图谱的各吸收峰位移与图1类似。Take 2g of N-isopropylacrylamide and 3mL of (γ-methacryloyl)propyltrimethoxysilane dissolved in 50ml of dioxane, add 100mg of benzoyl peroxide, blow nitrogen for 60min, and initiate at 65°C React for 24 hours. The solvent was rotary evaporated, dissolved in tetrahydrofuran, settled in ether, and dried; then dissolved in tetrahydrofuran, settled in petroleum ether, dried for later use, the molecular weight of the polymer was 41000, and the degree of dispersion was 3.0; the shifts of the absorption peaks in the nuclear magnetic spectrum were similar to those in Figure 1.

2.温度敏感水凝胶的合成2. Synthesis of temperature-sensitive hydrogels

取聚[N-异丙基丙烯酰胺-co-(γ-甲基丙烯酰基)丙基三甲氧基硅烷]100，200，500mg，加入1ml水，于室温下溶解，然后加入0.5ml磷酸盐/柠檬酸缓冲溶液，使反应液的pH值为6，反应于室温下进行72h，取出凝胶在水中浸泡除去杂质。凝胶的室温时的溶胀率分别为14，10，5。Take poly[N-isopropylacrylamide-co-(γ-methacryloyl)propyltrimethoxysilane] 100, 200, 500mg, add 1ml water, dissolve at room temperature, then add 0.5ml phosphate/ citric acid buffer solution, the pH value of the reaction solution was 6, the reaction was carried out at room temperature for 72 hours, and the gel was taken out and soaked in water to remove impurities. The swelling ratios of the gels at room temperature were 14, 10, and 5, respectively.

实施例3：Example 3:

取N-异丙基丙烯酰胺2g，和20ml(γ-甲基丙烯酰)丙基三甲氧基硅烷溶于50ml二氧六环，加入过氧化苯甲酰，通氮气除氧，于65℃下引发反应2-24小时。旋转蒸发溶剂，用四氢呋喃溶解，乙醚沉降，干燥；再用四氢呋喃溶解，石油醚沉降，(通过控制单体与引发剂过氧化苯甲酰的用量比和引发时间)可得到分子量在5000-100,000的聚合物；核磁图谱的各吸收峰位移与图1类似。Take 2g of N-isopropylacrylamide and 20ml of (γ-methacryloyl)propyltrimethoxysilane dissolved in 50ml of dioxane, add benzoyl peroxide, blow nitrogen to deoxygenate, at 65°C Initiate the reaction for 2-24 hours. Rotary evaporation of solvent, dissolution with tetrahydrofuran, precipitation of ether, drying; dissolution with tetrahydrofuran, precipitation of petroleum ether, (by controlling the amount ratio and initiation time of monomer and initiator benzoyl peroxide) can obtain molecular weight at 5000-100,000 Polymer; the displacement of each absorption peak in the NMR spectrum is similar to that in Figure 1.

2.温度敏感水凝胶的合成2. Synthesis of temperature-sensitive hydrogels

取聚[N-异丙基丙烯酰胺-co-(γ-甲基丙烯酰基)丙基三甲氧基硅烷]100mg，200mg，500mg，加入1ml水，于室温下溶解，然后加入0.5ml磷酸盐/柠檬酸缓冲溶液，使反应液的pH值为6，反应于室温下进行72h，得温度敏感水凝胶，取出凝胶在水中浸泡除去杂质。室温时所得凝胶的溶胀率在5-15之间。Take poly[N-isopropylacrylamide-co-(γ-methacryloyl)propyltrimethoxysilane] 100mg, 200mg, 500mg, add 1ml water, dissolve at room temperature, then add 0.5ml phosphate/ citric acid buffer solution, the pH value of the reaction solution is 6, and the reaction is carried out at room temperature for 72 hours to obtain a temperature-sensitive hydrogel, which is taken out and soaked in water to remove impurities. The resulting gel has a swelling ratio between 5 and 15 at room temperature.

实施例4：生物活性分子的负载和释放Example 4: Loading and Release of Bioactive Molecules

取实施例1得到的聚[N-异丙基丙烯酰胺-co-(γ-甲基丙烯酰基)丙基三甲氧基硅烷]200mg，小牛血清蛋白50mg，加入1ml水，于室温下溶解，然后加入0.5ml磷酸盐/柠檬酸缓冲溶液，使反应液的pH值为6，反应于室温下进行24h，取出凝胶在水中浸泡洗去小分子杂质。Take 200 mg of poly[N-isopropylacrylamide-co-(γ-methacryloyl)propyltrimethoxysilane] obtained in Example 1, 50 mg of bovine serum albumin, add 1 ml of water, and dissolve at room temperature. Then 0.5ml of phosphate/citric acid buffer solution was added to make the pH of the reaction solution 6, and the reaction was carried out at room temperature for 24 hours, and the gel was taken out and soaked in water to wash away small molecular impurities.

将以上负载小牛血清蛋白的凝胶放入20ml的水中，隔一定时间更换水溶液，同时测定小牛血清蛋白的释放量。图3为小牛血清蛋白的释放动力学，小牛血清蛋白分子(BSA)在46h内释放完全。从图4的蛋白质电泳，没有发现小牛血清蛋白任何性状的变化(图4中a为标准BSA、b为释放2小时、c为释放5.5小时、d为释放11小时、e为释放25.5小时的电泳图)。可见采用本发明控制释放生物大分子时，负载率高，可以很好地保持生物活性分子的活性。The above gel loaded with bovine serum albumin was put into 20ml of water, and the aqueous solution was replaced at regular intervals, and the released amount of bovine serum albumin was measured simultaneously. Fig. 3 is the release kinetics of bovine serum albumin, bovine serum albumin molecule (BSA) releases completely within 46h. From the protein electrophoresis of Fig. 4, do not find the change of any character of bovine serum albumin (in Fig. 4, a is standard BSA, b is that releases 2 hours, c is that releases 5.5 hours, d is that releases 11 hours, e is that releases 25.5 hours Electropherogram). It can be seen that when the present invention is used to control the release of biomacromolecules, the loading rate is high, and the activity of bioactive molecules can be well maintained.

实施例5：生物活性分子的负载和释放Example 5: Loading and Release of Bioactive Molecules

取实施例2得到的聚[N-异丙基丙烯酰胺-co-(γ-甲基丙烯酰基)丙基三甲氧基硅烷]100mg，小牛血清蛋白10或50mg，加入1ml水，于室温下溶解，然后加入0.5ml磷酸盐/柠檬酸缓冲溶液，使反应液的pH值为6，反应于室温下进行72h，取出凝胶在水中浸泡洗去小分子杂质。Take 100 mg of poly[N-isopropylacrylamide-co-(γ-methacryloyl) propyltrimethoxysilane] obtained in Example 2, 10 or 50 mg of bovine serum albumin, add 1 ml of water, and Dissolve, then add 0.5ml phosphate/citric acid buffer solution to make the pH of the reaction solution 6, react at room temperature for 72h, take out the gel and soak it in water to wash away small molecular impurities.

将以上负载小牛血清蛋白的凝胶放入20ml的水中，隔一定时间更换水溶液，同时测定小牛血清蛋白的释放量。小牛血清蛋白在9、28小时内释放完全。The above gel loaded with bovine serum albumin was put into 20ml of water, and the aqueous solution was replaced at regular intervals, and the released amount of bovine serum albumin was measured simultaneously. The bovine serum albumin is completely released within 9 and 28 hours.

Claims

Translated fromChinese

1.含硅氧烷侧链的温度敏感线性聚合物，其特征在于，由下法制得：按99g∶1ml-1g∶50ml的比例取N-异丙基丙烯酰胺和(γ-甲基丙烯酰基)丙基三甲氧基硅烷作为单体，并按1g∶1-10ml的比例将单体溶于1，4-二氧六环，以过氧化苯甲酰为引发剂在65-90℃引发聚合反应2-24小时，得到分子量为5000-100,000的含硅氧烷侧链的温度敏感线性聚合物。1. The temperature-sensitive linear polymer containing siloxane side chain is characterized in that it is prepared by the following method: take N-isopropylacrylamide and (γ-methacryloyl) in the ratio of 99g: 1ml-1g: 50ml ) Propyltrimethoxysilane as a monomer, and dissolve the monomer in 1,4-dioxane at a ratio of 1g: 1-10ml, and use benzoyl peroxide as an initiator to initiate polymerization at 65-90°C After reacting for 2-24 hours, a temperature-sensitive linear polymer containing siloxane side chains with a molecular weight of 5000-100,000 is obtained.

2.权利要求1所述的聚合物的合成方法，其特征在于：按99g∶1ml-1g∶50ml的比例取N-异丙基丙烯酰胺和(γ-甲基丙烯酰基)丙基三甲氧基硅烷作为单体，并按1g∶1-10ml的比例将单体溶于1，4-二氧六环，以过氧化苯甲酰为引发剂在65-90℃引发聚合反应2-24小时，得到分子量为5000-100,000的含硅氧烷侧链的温度敏感线性聚合物。2. the synthetic method of polymer described in claim 1 is characterized in that: get N-isopropylacrylamide and (γ-methacryloyl) propyl trimethoxyl group by the ratio of 99g: 1ml-1g: 50ml Silane is used as a monomer, and the monomer is dissolved in 1,4-dioxane at a ratio of 1g: 1-10ml, and the polymerization reaction is initiated at 65-90°C for 2-24 hours with benzoyl peroxide as an initiator. A temperature-sensitive linear polymer containing siloxane side chains with a molecular weight of 5000-100,000 was obtained.

3.根据权利要求2所述的制备方法，其特征在于：单体与引发剂的用量重量比为100∶0.2-5。3. The preparation method according to claim 2, characterized in that: the weight ratio of monomer to initiator is 100:0.2-5.

4.温度敏感水凝胶，其特征在于，由下法制得：将权利要求1所述含硅氧烷侧链的温度敏感线性聚合物溶于水制得浓度为5-60wt％的溶液，然后加入磷酸盐/柠檬酸盐的混合溶液至溶液的pH值为3-12，水解交联反应12-72小时，得到温度敏感水凝胶。4. The temperature-sensitive hydrogel is characterized in that, it is obtained by the following method: the temperature-sensitive linear polymer containing siloxane side chains described in claim 1 is dissolved in water to obtain a solution with a concentration of 5-60wt%, and then The mixed solution of phosphate/citrate is added until the pH value of the solution is 3-12, and the hydrolysis and cross-linking reaction is carried out for 12-72 hours to obtain a temperature-sensitive hydrogel.

5.权利要求4所述的温度敏感水凝胶的制备方法，其特征是：将权利要求I所述含硅氧烷侧链的温度敏感线性聚合物溶于水制得浓度为5-60wt％的溶液，然后加入磷酸盐/柠檬酸盐的混合溶液至溶液的pH值为3-12，水解交联反应12-72小时，得到温度敏感水凝胶。5. the preparation method of temperature-sensitive hydrogel described in claim 4 is characterized in that: the temperature-sensitive linear polymer containing siloxane side chain described in claim 1 is dissolved in water and makes concentration be 5-60wt% solution, and then add a mixed solution of phosphate/citrate until the pH value of the solution is 3-12, and perform a hydrolysis and cross-linking reaction for 12-72 hours to obtain a temperature-sensitive hydrogel.

6.权利要求4所述的温度敏感水凝胶在生物活性分子的负载、固定化或控制释放中的应用。6. The application of the temperature-sensitive hydrogel according to claim 4 in the loading, immobilization or controlled release of bioactive molecules.