Kumar Pawan, Truong Lynda, Baker Ysobel Ruth, El-Sagheer Afaf Helmy, Brown Tom
Department of Chemistry, University of Oxford, 12 Mansfield Road, Oxford OX1 3TA, U.K.
Chemistry Branch, Department of Science and Mathematics, Faculty of Petroleum and Mining Engineering, Suez University, Suez 43721, Egypt.
ACS Omega. 2018 Jun 30;3(6):6976-6987. doi: 10.1021/acsomega.8b01086. Epub 2018 Jun 27.
Dinucleoside phosphoramidites containing a triazole internucleotide linkage flanked by locked nucleic acid (LNA) were synthesized and incorporated into oligonucleotides (ONs). ONs bearing both LNA and triazole at multiple sites were obtained and their biophysical properties including enzymatic stability and binding affinity for RNA and DNA targets were studied. t-LNAs with four incorporations of a dinucleoside monomer having LNA on either side of the triazole linkage bind to their RNA target with significantly higher affinity and greater specificity than unmodified oligonucleotides, and are remarkably stable to nuclease degradation. A similar but reduced effect on enzymatic stability and binding affinity was noted for LNA only on the 3'-side of the triazole linkage. Thus, by combining unnatural triazole linkages and LNA in one unit (t-LNA), we produced a promising class of ONs with reduced anionic charge and potential for antisense applications.
合成了含有由锁核酸（LNA）侧翼的三唑核苷酸间连接的二核苷亚磷酰胺，并将其掺入寡核苷酸（ONs）中。获得了在多个位点同时带有LNA和三唑的ONs，并研究了它们的生物物理性质，包括酶稳定性以及对RNA和DNA靶标的结合亲和力。在三唑连接两侧均具有LNA的二核苷单体进行四次掺入的t-LNA与它们的RNA靶标的结合亲和力明显高于未修饰的寡核苷酸，且具有更高的特异性，并且对核酸酶降解具有显著的稳定性。对于仅在三唑连接的3'侧具有LNA的情况，观察到对酶稳定性和结合亲和力有类似但减弱的影响。因此，通过将非天然的三唑连接和LNA结合在一个单元（t-LNA）中，我们制备了一类有前景的ONs，其阴离子电荷减少，具有反义应用潜力。
