Synthesis of stable azide and alkyne functionalized phosphoramidite nucleosides

Tetrahedron Lett. 2019 Jan 17;60(3):211-213. doi: 10.1016/j.tetlet.2018.12.018. Epub 2018 Dec 8.


The use of CuAAC chemistry to crosslink and stabilize oligonucleotides has been limited by the incompatibility of azides with the phosphoramidites used in automated oligonucleotide synthesis. Herein we report optimized reaction conditions to synthesize azide derivatives of thymidine and cytidine phosphoramidites. Investigation of the stability of the novel phosphoramidites using 31P NMR at room temperature showed less than 10% degradation after 6 hours. The azide modified thymidine was successfully utilized as an internal modifier in the standard phosphoramidite synthesis of a DNA sequence. The synthesized azide and alkyne derivatives of pyrimidines will allow efficient incorporation of azide and alkyne click pairs into nucleic acids, thus widening the applicability of click chemistry in investigating the chemistry of nucleic acids.

Keywords: Chemical biology; Click Chemistry; CuAAC; Oligonucleotide synthesis; Phosphoramidites.