Varizhuk Anna M, Zatsepin Timofei S, Golovin Andrey V, Belyaev Evgeny S, Kostyukevich Yury I, Dedkov Vladimir G, Shipulin German A, Shpakovski George V, Aralov Andrey V
Research and Clinical Center for Physical Chemical Medicine, 119435 Moscow, Russia; Department of Structure-Functional Analysis of Biopolymers, Engelhardt Institute of Molecular Biology, Vavilov Str. 32, Moscow 119991, Russia.
Chemistry Department, Lomonosov Moscow State University, Leninskie Gory, Moscow 119991, Russia; Central Research Institute of Epidemiology, Novogireevskaya 3a, Moscow 111123, Russia; Skolkovo Institute of Science and Technology, 3 Nobel Street, Skolkovo, Moscow Region 143026, Russia.
Bioorg Med Chem. 2017 Jul 15;25(14):3597-3605. doi: 10.1016/j.bmc.2017.03.062. Epub 2017 Mar 31.
Nowadays modified oligonucleotides are widely used in diagnostics and as novel therapeutics. Introduction of modified or unnatural residues into oligonucleotides allows fine tuning of their binding properties to complementary nucleic acids and leads to improved stability both in vitro and in vivo. Previously it was demonstrated that insertion of phenoxazine nucleotides with various groups in C9-position into oligonucleotides leads to a significant increase of duplex stability with complementary DNA and RNA. Here the synthesis of a novel G-clamp nucleoside analogue (G-clamp) bearing 2-aminoethyl tether at C8-atom is presented. Introduction of such modified residues into oligonucleotides lead to enhanced specificity of duplex formation towards complementary DNA and RNA targets with increased thermal and 3'-exonuclease stability. According to CD-spectroscopy studies G-clamp does not substantially disrupt helix geometry. Primers containing G-clamp demonstrated superior sensitivity in qPCR detection of dsRNA of Kemerovo virus in comparison to native oligonucleotides.
如今，修饰的寡核苷酸广泛应用于诊断领域，并作为新型治疗药物。在寡核苷酸中引入修饰的或非天然的残基，可以微调它们与互补核酸的结合特性，并提高其在体外和体内的稳定性。此前有研究表明，将在C9位带有不同基团的吩恶嗪核苷酸插入寡核苷酸中，会显著提高与互补DNA和RNA形成的双链体的稳定性。本文介绍了一种在C8原子上带有2-氨基乙基连接链的新型G-钳核苷类似物（G-钳）的合成。将这种修饰的残基引入寡核苷酸中，可增强与互补DNA和RNA靶标形成双链体的特异性，并提高热稳定性和3'-外切核酸酶稳定性。根据圆二色光谱研究，G-钳不会显著破坏螺旋结构。与天然寡核苷酸相比，含有G-钳的引物在克麦罗沃病毒双链RNA的qPCR检测中表现出更高的灵敏度。
