摘要
通过将开环易位聚合反应和CuAAC反应联用制备了一系列含糖(共)聚合物.首先采用Cu(I)催化的叠氮-端炔[3+2]环加成(CuAAC)合成了含无保护基团的α-D-甘露糖、β-D-葡萄糖和β-D-半乳糖的7-氧杂降冰片烯衍生物单体;接着利用Grubbs三代催化剂在常温常压下的均相有机溶剂中对不同类的含糖单体进行开环易位聚合(ROMP),通过改变含糖单体的种类和比例,得到了一系列结构明确的含糖均聚和共聚物P1~P11.用核磁共振谱(NMR)和高分辨质谱(HRMS)对合成的糖单体的结构及分子量进行表征.含糖聚合物的分子量(分布)及结构通过凝胶渗透色谱仪(GPC)和核磁共振谱(NMR)进行表征,结果表明分子量可控(Mn=1.3×10~4~2.7×10~4),分子量分布较窄(PDI=1.22~1.45).进一步采用浊度法、动态光散射和等温滴定量热仪研究了含糖聚合物与刀豆蛋白A (concanavalin A,Con A)的特异性识别.浊度法研究发现,共聚物α-D-甘露糖的比例越大,其与Con A的特异性识别能力越强,而只含β-D-半乳糖P9或β-D-葡萄糖P5对刀豆蛋白没有特异性识别.动态光散射实验证实,随着刀豆蛋白A的加入,含甘露糖的溶液中的粒径明显增大,含糖聚合物溶液的粒径由100 nm左右增加到1000 nm左右,而不含甘露糖的聚合物几乎没有变化.等温滴定量热仪测定3种代表性共聚物与Con A的结合常数Ka分别为P3 (50 mol%α-D-甘露糖:50 mol%β-D-葡萄糖,K_a=1.58×10~6 L/mol),P7 (50 mol%α-D-甘露糖:50 mol%β-D-半乳糖,K_a=2.23×10~6 L/mol)和P11 (50 mol%α-D-甘露糖:50 mol%非糖基团,K_a=2.05×10~5 L/mol).可以看出P11与Con A结合能力相对于P3和P7要小很多,说明β-D-葡萄糖和β-D-半乳糖对α-D-甘露糖与Con A的识别作用有较强的协同效应.
A series of glycopolymers were prepared through combining ring-opening metathesis polymerization(ROMP) and CuAAC reaction.Firstly,a wide range of exo-7-oxanorbornene derivative glycomonomers without protecting groups were synthesized via a copper(I)-catalyzed azide-alkyne Huisgen cycloaddition(CuAAC)reaction,including α-D-mannose,β-D-glucose,and β-D-galactose.A series of well-defined glycopolymers were then obtained from various types and proportions of the above glycomonomers using ring-opening metathesis polymerization(ROMP) with the 3rd Grubbs catalyst in homogeneous organic solvent.Molecular weight and polydispersity index(PDI) of the glycopolymers were characterized by NMR spectroscopy and GPC,from which the well-controlled molecular weight(M_n = 1.3 × 10~4-2.7 × 10~4) in narrow distribution(PDI = 1.22 ~ 1.45) was confirmed.Turbidity measurement,dynamic light scattering(DLS),and isothermal titration calorimetry(ITC)were carried out to investigate the specific recognition of glycopolymers with concanavalin A(Con A).Turbidimetric study suggested a stronger binding ability of glycopolymers with Con A at higher ratio of α-Dmannose in glycopolymers.In comparison,those composed solely of β-D-galactose(P9) or β-D-glucose(P5)could not bind to Con A.Dynamic light scattering experiments demonstrated that the particle sizes of glycopolymers containing α-D-mannose approached 1000 nm with the addition of Con A(originally 100 nm),while the glycopolymers without α-D-mannose showed little size variation.Binding constants(K_a) of the three glycopolymers P3(50 mol% α-D-mannose,50 mol% β-D-glucose),P7(50 mol% α-D-mannose,50 mol% β-Dgalactose),and P11(50 mol% α-D-mannose,50 mol% non-sugar motif) with Con A were 1.58 × 10~6,2.23 × 10~6,and 2.05 × 10~5 L/mol,respectively,as measured by isothermal titration calorimetry.P11 exhibited much weaker ability to bind with Con A than P3 and P7 did,which implied a synergistic effect of β-D-glucose and β-Dgalactose on the recognition of α-D-mannose with Con A.
引文
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