Biomolecular Nanoscale Engineering Center, Department of Physics, Chemistry and Pharmacy, University of Southern Denmark, Campusvej 55, 5230, Odense M, Denmark.
Department of Chemistry, Biomolecular Nanoscale Engineering Center, University of Copenhagen, Thorvaldsensvej 40, Frederiksberg, 1871, Denmark.
Chemistry. 2021 Jan 18;27(4):1416-1422. doi: 10.1002/chem.202004495. Epub 2020 Dec 16.
Attachment of cationic moieties to oligonucleotides (ONs) promises not only to increase the binding affinity of antisense ONs by reducing charge repulsion between the two negatively charged strands of a duplex, but also to augment their in vivo stability against nucleases. In this study, polyamine functionality was introduced into ONs by means of 2'-amino-LNA scaffolds. The resulting ONs exhibited efficient binding towards ssDNA, ssRNA and dsDNA targets, and the 2'-amino-LNA analogue carrying a triaminated linker showed the most pronounced duplex- and triplex-stabilizing effect. Molecular modelling revealed that favourable conformational and electrostatic effects led to salt-bridge formation between positively charged polyamine moieties and the Watson-Hoogsteen groove of the dsDNA targets, resulting in the observed triplex stabilization. All the investigated monomers showed increased resistance against 3'-nucleolytic digestion relative to the non-functionalized controls.
将阳离子部分连接到寡核苷酸 (ONs) 上不仅有望通过减少双链体中两条带负电荷的链之间的电荷排斥来增加反义 ONs 的结合亲和力，而且还可以增强它们对核酸酶的体内稳定性。在这项研究中，通过 2'-氨基-LNA 支架将聚胺功能引入到 ONs 中。所得的 ONs 对 ssDNA、ssRNA 和 dsDNA 靶标表现出有效的结合，并且携带三氨基连接子的 2'-氨基-LNA 类似物表现出最显著的双链体和三聚体稳定作用。分子建模表明，有利的构象和静电效应导致带正电荷的聚胺部分与 dsDNA 靶标的 Watson-Hoogsteen 沟之间形成盐桥，从而导致观察到的三聚体稳定。与非功能化对照相比，所有研究的单体对 3'-核酸酶消化的抗性均有所提高。
