Preparation of Glucose-Responsive Phenylboronic Acid-Containing Polymers and Their Application to Artificial Pancreas

葡萄糖响应性含苯基硼酸聚合物的制备及其在人工胰腺中的应用

• 批准号: 07559020
• 负责人: SAKURAI Yasuhisa
• 金额: $ 3.33万
• 依托单位: Tokyo Women's Medical College
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07559020/
• 关键词: Phenylboronic acid; Glucose; Insulin delivery; Artificial pancreas; Polymer complex; intelligent materials; Physiological pH; グルコース応答性; ドラッグデリバリー; ポリビニルアルコール; 高分子間コンプレックス; センサー; 膨潤・収縮; イオン伝導度

A new "Intelligent" polymer system sensitive to glucose concentration have been studied as a candidate material for chemically regulated insulin release system. Phenylboronic acid is capable to form reversible binding with cis-diol substances. Glucose responsive insulin release system has been studied with utilization of phenylboronic acid polymers for the key material to exchange reaction between gluconic acid modified insulin (G-Ins) and glucose molecule. CompIexation of various diol-containing molecules with PBA gel beads [poly (3-methacrylamidophenylboronic acid-co-acrylamide-co-N,N'-methylenebis- (acrylamide)] was evaluated using frontal chromatography at pH 8.5. The structural features of the diol-containing molecules strongly influenced their binding to PBA gel beads. In particular, open-chain mono saccharides demonstrated higher association constants (ca. 9.5X10X^2 to 5.1X10X^3 l/mol) than glucose (ca. 6.3X10X^2 l/mol). G-Ins was synthesized, and was bound onto PBA gel column. … More Then, the G-Ins release profile in response to varying glucose concentrations was investigated at pH 8.5. The released concentration of G-Ins from the polymer was pulsatile in response to the repeated stepwise concentration changes of glucose. Other remarkable results are that the linearity between glucose concentration and peak height of released G-lns, and that the release response have no lag time to change of glucose concentration. To enhance binding between PBA and glucose under physiological pH 7.4, an amine component was incorporated into the polymer chain along with PBA [poly (3-methacrylamidophenylboronic acid-co-N,N-dimethylaminopropylacrylamide-co-acrylamide-co-N,N'-methylenebis (acrylamide)]. The proportion of the amount of PBA groups complexed with glucose versus total amount of PBA groups was determined by batch method. Compared to PBA copolymer synthesized without the amine component, the proportion increased as a function of the amine content and the pH of the buffer. These results confirm that the interaction of neighboring amines (unprotonated) with PBA strengthens the binding with glucose, especially at pH 7.4 and above. Release property of G-Ins from the amine-containing PBA gel beads at pH 7.4 was evaluated using a liquid chromatography system. Amine-containing PBA gel showed less rapid decrease of G-Ins in the column under buffer-rinse over PBA gel. The released amount of G-Ins from amine-containing PBA gel is much greater than for the PBA gel, responding to the addition of glucose at physiological pH.The results from long-term release studies demonstrate that the amine-containing PBA gel has the capability to respond to glucose challenge over 120 hours. Furthermore, a linearity of released G-Ins was observed with glucose concentration. These results demonstrate the feasibility of an insulin-responsive system to achieve insulin release in response to glucose under physiological conditions. Less

研究了一种对葡萄糖浓度敏感的新型“智能”聚合物体系，作为化学调节胰岛素释放系统的候选材料。苯基硼酸能够与顺式二醇物质形成可逆结合。利用苯硼酸聚合物作为葡萄糖酸修饰胰岛素（G-Ins）与葡萄糖分子交换反应的关键材料，研究了葡萄糖响应性胰岛素释放系统。在pH 8.5下使用前沿色谱法评价各种含二醇的分子与PBA凝胶珠[聚（3-甲基丙烯酰胺基苯基硼酸-共-丙烯酰胺-共-N，N '-亚甲基双-（丙烯酰胺）]的复合。含二醇分子的结构特征强烈影响其与PBA凝胶珠的结合。特别是，开链单核苷酸表现出更高的缔合常数（约。9.5X10X^2至5.1X10X^3 l/mol）比葡萄糖（约6.3X10X^2 l/mol）。合成了G-Ins，并将其结合到PBA凝胶柱上。 ...更多信息 然后，响应于不同的葡萄糖浓度的G-Ins释放曲线进行了研究，在pH 8.5。从聚合物中释放的G-Ins的浓度是脉动的响应于葡萄糖的重复逐步浓度变化。葡萄糖浓度与释放峰高呈线性关系，释放响应对葡萄糖浓度的变化无滞后。为了在生理pH 7.4下增强PBA和葡萄糖之间的结合，将胺组分与PBA [聚（3-甲基丙烯酰胺基苯基硼酸-共-N，N-二甲基氨基丙基丙烯酰胺-共-丙烯酰胺-共-N，N '-亚甲基双（丙烯酰胺）]一起沿着掺入聚合物链中。通过分批法测定与葡萄糖复合的PBA基团的量相对于PBA基团的总量的比例。与不含胺组分合成的PBA共聚物相比，该比例作为胺含量和缓冲液pH的函数而增加。这些结果证实了邻近胺（未质子化的）与PBA的相互作用增强了与葡萄糖的结合，特别是在pH 7.4及以上。使用液相色谱系统评价在pH 7.4下G-Ins从含胺PBA凝胶珠的释放性质。含胺的PBA凝胶在PBA凝胶上的缓冲液冲洗下显示出柱中G-Ins的较不快速的减少。从含胺PBA凝胶中释放的G-Ins的量远大于PBA凝胶，响应于在生理pH下加入葡萄糖。长期释放研究的结果表明，含胺PBA凝胶具有响应葡萄糖挑战超过120小时的能力。此外，观察到释放的G-Ins与葡萄糖浓度呈线性关系。这些结果证明了胰岛素响应系统在生理条件下响应于葡萄糖而实现胰岛素释放的可行性。少

项目成果

Takeshi lkeya, Hiroaki Miyazaki, Kazunori Kataoka, Teruo Okano, Yasuhisa Sakurai: "Activation of lymphocytes by poly (N-vinylpyrrolidon-alt-maleic acid) derivatized with phenylboronic acid" Journal of Japanese Society for Biomaterials. 14. 260-266 (1996)

Takeshi lkeya、Hiroaki Miyazaki、Kazunori Kataoka、Teruo Okano、Yasuhisa Sakurai：“用苯基硼酸衍生的聚（N-乙烯基吡咯烷酮-alt-马来酸）激活淋巴细胞”日本生物材料学会杂志。

D.Shiino: "Amine containing phenylboronic acid gels for glucose-responsive insulin release under physiological pH" Journal of Controlled Release. 37. 269-276 (1995)

D.Shiino：“含胺苯基硼酸凝胶，用于在生理 pH 值下释放葡萄糖响应性胰岛素”《控释杂志》。

Daijiro Shiino, Yoshishige Murata, Akira Kubo, Yong Joo Kim, Kazunori Kataoka, Yoshiyuki Koyama, Akihiko Kikuchi, Masayuki Yokoyama, Yasuhisa Sakurai, Teruo Okano: "Amine containing phenylboronic acid gel for glucose-responsive insulin release under physi

Daijiro Shiino、Yoshishige Murata、Akira Kubo、Yong Joo Kim、Kazunori Kataoka、Yoshiyuki Koyama、Akihiko Kikuchi、Masayuki Yokoyama、Yasuhisa Sakurai、Teruo Okano：“含胺苯硼酸凝胶，用于物理条件下葡萄糖响应性胰岛素释放

I.Hisamitsu: "Glucose-responsive gel from pheylborate polymer and poly(viny alchol):prompt response at physiological pH through the interaction of borate with amino group in the gel" Pharmaceutical Research. 14. 289-293 (1997)

I.Hisamitsu：“由苯硼酸盐聚合物和聚（乙烯醇）制成的葡萄糖响应凝胶：通过硼酸盐与凝胶中氨基的相互作用在生理 pH 下迅速响应”药物研究。

Kazunori Kataoka, Issei Hisamitsu, Naoko Sayama, Teruo Okano, Yasuhisa Sakurai: "Novel sensing for glucose based on the complex formation between phenylborate and fluorescent diol compounds" Journal of Biochemistry. 117. 1145-1147 (1995)

Kazunori Kataoka、Issei Hisamitsu、Naoko Sayama、Teruo Okano、Yasuhisa Sakurai：“基于硼酸苯酯和荧光二醇化合物之间的复合物形成的新型葡萄糖传感”《生物化学杂志》。