Increased Affinity of 2′-O-(2-Methoxyethyl)-Modified Oligonucleotides to RNA through Conjugation of Spermine at Cytidines

Structural modification at the 2′-O-position of riboses in oligonucleotide therapeutics is of critical importance for their use as drugs. To date, the methoxyethyl (MOE) substituent is the most important and features in dozens of antisense oligonucleotides that have been tested in clinical trials. Yet, the search for new improved modifications continues in a quest for increased oligonucleotide potency, improved transport in vivo and favorable metabolism. Recently, we described how the conjugation of spermine groups to pyrimidines in oligonucleotides vastly increases their affinity for complementary RNAs through accelerated binding kinetics. Here we describe how spermines can be linked to the exocyclic amino groups of cytidines in MOE-oligonucleotides employing a straightforward ‘convertible nucleoside approach’ during solid phase synthesis. Singly- or doubly-modified oligonucleotides show greatly enhanced affinity for complementary RNA, with potential for a new generation of MOE-based oligonucleotide drugs.