Design of supramolecular ligands that can multivalently interact with receptor proteins on cellular membranes

与细胞膜上受体蛋白多价相互作用的超分子配体的设计

• 批准号: 14380397
• 负责人: YUI Nobuhiko
• 金额: $ 9.15万
• 依托单位: Japan Advanced Institute of Science and Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-14380397/
• 关键词: Polyrotaxane; Cyclodextrins; Poly(ethylene glycol); Supramolecular; Sccharides; Gene; Gene expression; Biomaterials; シクロデキストリン; 多価相互作用; 軟骨細胞; 骨芽細胞; マルトース; 分子運動性; 分解制御; コンカナバリンA; トランスポーター; 赤血球凝集阻害

In this research, new biomaterials using a polyrotaxane was designed and examined to achieve strong binding with receptor proteins on cellular membranes. The most definitive feature in polyrotaxanes, in which many cyclic compounds are threaded onto linear polymeric chains capped with bulky end-groups, is the noncovalent bonds between the cyclic compounds and the polymeric chain. Through the research, we found and clarified the following matters.Multivalent Molecular Recognition Based on Sliding Motion of Saccharide-Polyrotaxane Conjugates : Saccharide, one of the ligands for targeting, in saccharide-polyrotaxane conjugates could easily diffuse in buffers via sliding and rotational motion of α-cyclodextrins (α-CDs) along a poly(ethylene glycol) (PEG) chain. A maltose-polyrotaxane conjugate with appropriate threading number of α-CDs exhibits high mobility of maltose in aqueous conditions. A high mobility of saccharide groups and α-Cds was proved by spin-lattice relaxation time (T_1) and … More spin-spin relaxation time (T_2) analysis. The association constant (K_a) between the conjugate and Concanavalin A (Con A) was in the range of 6 powers. The features of high mobility of the ligands and preserving the water clusters led to gaining a large negative enthalpy.Erosion Time-Controllable Hydrogels Using Polyrotaxanes for Cartilege Tissue Enginnering : Hydrolyzable polyrotaxanes were utilized as a cross-linker for preparing PEG-polyrotaxane hydrogels (PEG-PRX gels) for biomedical applications. The hydrolysis of our designed PEG-PRX gels was found to well controlled by inclusion complexation of ester groups located at the terminal of the PEG in the polyrotaxanes, and the time to complete hydrogel erosion varies from a few days to more than a half year in spite of their highly swollen states in physiological conditions. By using the degradation control, the PEG-PRX gels could be designed as scaffolds for cartilege cultivation.Plasmid DNA Delivery to Nucleus Based on Supramolecular Characteristics of Biocleavable Polyrotaxane : A biocleavable polyrotaxane, having a necklace-like structure consisting of many cationic α-CDs and a disulfide-introduced PEG, was examined as a non-viral gene carrier. The polyrotaxane formed a stable polyplex having positively charged surface even at low charge ratio. The pDNA decondensation occurred through disulfide cleavage of the polyrotaxane and subsequent supramolecular dissociation of the non-covalent linkages between α-CDs and PEG. Rapid endosomal escape was observed, and the transfection of the DMAE-SS-PRX polyplex is independent of the amount of free polycation. Those properties played a key role for delivery of pDNA clusters to nucleus Less

在这项研究中，设计并检查了使用聚轮烷的新生物材料，以实现与细胞膜上受体蛋白的强结合。在聚轮烷中，其中许多环状化合物被穿线到用大体积端基封端的线性聚合物链上，最确定的特征是环状化合物和聚合物链之间的非共价键。通过研究，我们发现并澄清了以下问题：基于糖-聚轮烷缀合物滑动运动的多价分子识别：糖-聚轮烷缀合物中的糖是靶向配体之一，可以通过α-环糊精（α-CD）沿着聚乙二醇（PEG）链的滑动和旋转运动在缓冲液中轻松扩散。具有适当的α-CD螺纹数的麦芽糖-聚轮烷缀合物在水性条件下表现出麦芽糖的高迁移率。自旋-晶格弛豫时间（T_1）证明了糖基和α-Cds的高迁移率， ...更多信息 自旋-自旋弛豫时间（T_2）分析。偶联物与伴刀豆球蛋白A（ConA）的结合常数（Ka）在6次方范围内。高流动性的配体和保存的水簇的特点，导致获得一个大的负焓。腐蚀时间可控的水凝胶使用聚轮烷用于软骨组织工程：可水解的聚轮烷作为交联剂，用于制备PEG-聚轮烷水凝胶（PEG-PRX凝胶）的生物医学应用。我们设计的PEG-PRX凝胶的水解被发现通过位于聚轮烷中PEG末端的酯基的包合络合来很好地控制，并且完成水凝胶侵蚀的时间从几天到半年以上不等，尽管它们在生理条件下处于高度溶胀状态。基于生物可切割聚轮烷超分子特性的质粒DNA向细胞核的递送：生物可切割聚轮烷具有由许多阳离子α-CD和引入二硫键的PEG组成的项链状结构，可作为非病毒基因载体。该聚轮烷即使在低电荷比下也形成具有带正电荷的表面的稳定的聚合复合物。通过聚轮烷的二硫键裂解和随后α-CD和PEG之间的非共价键的超分子解离发生pDNA解凝聚。观察到内体快速逃逸，并且DMAE-SS-PRX多聚物的转染与游离多聚阳离子的量无关。这些性质在将pDNA簇递送到细胞核中起着关键作用。

项目成果

Temperature-controlled erosion of poly(N-isopropylacrylamide)-based hydrogels crosslinked by methacrylate-introduced hydrolyzable polyrotaxane

甲基丙烯酸酯引入的可水解聚轮烷交联的聚（N-异丙基丙烯酰胺）基水凝胶的温控侵蚀

• 发表时间: 2005
• 期刊: Science and Technology of Advanced Materials 6
• 作者: T.Ooya;M.Akutsu;Y.Kumashiro;N.Yui
• 通讯作者: N.Yui

T.Ichi, T.Ooya, N.Yui: "Supramolecular control of ester hydrolysis in poly(ethylene glycol) interlocked hydrogels"Macomol.Biosci.. 3(7). 373-380 (2003)

T.Ichi、T.Ooya、N.Yui：“聚乙二醇联锁水凝胶中酯水解的超分子控制”Macomol.Biosci.. 3(7)。

Effects of polyrotaxane structure on polyion complexation with DNA

• DOI: 10.1016/j.stam.2003.12.014
• 发表时间: 2004-05-01
• 期刊: SCIENCE AND TECHNOLOGY OF ADVANCED MATERIALS
• 影响因子: 5.5
• 作者: Ooya, T;Yamashita, A;Yui, N
• 通讯作者: Yui, N

M.Eguchi, T.Ooya, N.Yui: "Controlling the mechanism of trypsin inhibition by the numbers of α-cyclodextrins and carboxyl groups in carboxyethylester-polyrotaxanes"J.Controlled Release. (印刷中). (2004)

M.Eguchi、T.Ooya、N.Yui：“通过羧乙酯-聚轮烷中的 α-环糊精和羧基数量控制胰蛋白酶抑制的机制”J.Controlled Release（印刷中）。

N.Yui, T.Ooya: "Polyrotaxanes : challenge to multivalent binding with biological receptors on cell surfaces"Mater.Sci.Forum. 426-432. 3243-3248 (2003)

N.Yui、T.Ooya：“聚轮烷：对细胞表面生物受体多价结合的挑战”Mater.Sci.Forum。