Chemistry Department M.V. Lomonosov Moscow State University, Leninskie gory 1-3, Moscow, 119991 Russian Federation, Russia.
Curr Med Chem. 2012;19(30):5232-7. doi: 10.2174/092986712803530575.
G-quadruplex-based DNA aptamers for human thrombin represent promising pharmaceuticals having high anticoagulant activity, rapid clearance from the bloodstream, and availability of DNA antidote. High specificity of aptamers minimizes side-effects but at the same time leads to difficulties when choosing animal model for preclinical trials. Well described minimal G-quadruplex 15-TBA, elongated with hinge and duplex regions 31-TBA, and developed recently modular aptamer RA-36 were investigated thoroughly and compared in this paper. To estimate the species-specificity of aptamers the inhibition constants and types were determined for human fibrinogen hydrolysis with human, bovine, porcine, rabbit, rat, and mouse thrombins using turbidimetric assay. Coagulation tests on human fibrinogen, factor II deficient human plasma and animal plasmas were conducted to verify turbidimetric data and reveal either the concentration effects or the influence of other plasma components. And finally, a set of tests on blood clotting cascade was performed to investigate target-specificity of RA-36 aptamer.
基于 G-四链体的人凝血酶 DNA 适体具有高抗凝活性、快速从血液中清除以及可提供 DNA 解毒剂等特点，是很有前途的药物。适体的高特异性可最大程度地减少副作用，但同时在选择临床前试验的动物模型时也会带来困难。本文深入研究并比较了描述良好的最小 G-四链体 15-TBA、带有铰链和双链区 31-TBA 的伸长体以及最近开发的模块化适体 RA-36。为了评估适体的物种特异性，使用比浊法测定了人凝血酶、牛凝血酶、猪凝血酶、兔凝血酶、鼠凝血酶和兔凝血酶对人纤维蛋白原水解的抑制常数和类型。进行了人纤维蛋白原、因子 II 缺乏的人血浆和动物血浆的凝血试验，以验证比浊数据，并揭示浓度效应或其他血浆成分的影响。最后，进行了一组血液凝固级联试验，以研究 RA-36 适体的靶特异性。
