A Versatile Solid-Phase Approach to the Synthesis of Oligonucleotide Conjugates with Biodegradable Hydrazone Linker

One of the ways to efficiently deliver various drugs, including therapeutic nucleic acids, into the cells is conjugating them with different transport ligands via labile or stable bonds. A convenient solid-phase approach for the synthesis of 5 '-conjugates of oligonucleotides with biodegradable pH-sensitive hydrazone covalent bonds is proposed in this article. The approach relies on introducing a hydrazide of the ligand under aqueous/organic media to a fully protected support-bound oligonucleotide containing aldehyde function at the 5 '-end. We demonstrated the proof-of-principle of this approach by synthesizing 5 '-lipophilic (e.g., cholesterol and alpha-tocopherol) conjugates of modified siRNA and non-coding RNAs imported into mitochondria (antireplicative RNAs and guide RNAs for Mito-CRISPR/system). The developed method has the potential to be extended for the synthesis of pH-sensitive conjugates of oligonucleotides of different types (ribo-, deoxyribo-, 2 '-O-methylribo-, and others) with ligands of different nature.

Automated summary

A convenient solid-phase approach for synthesizing 5'-conjugates of oligonucleotides with biodegradable pH-sensitive hydrazone covalent bonds is proposed in this study. The proof-of-principle of this approach was demonstrated by synthesizing lipophilic conjugates of modified siRNA and non-coding RNAs imported into mitochondria. This method has the potential to be extended for synthesizing pH-sensitive conjugates of oligonucleotides with ligands of different nature.