Structural Basis of Duplex Thermodynamic Stability and Enhanced Nuclease Resistance of 5 %26apos;-C-Methyl Pyrimidine-Modified Oligonucleotides

作者：Kel' in Alexander V; Zlatev Ivan; Harp Joel; Jayaraman Muthusamy; Bi**e Anna; O' Shea Jonathan; Taneja Nate; Manoharan Rajar M; Khan Saeed; Charisse Klaus; Maier Martin A; Egli Martin; Rajeev Kallanthottathil G; Manoharan Muthiah

来源：Journal of Organic Chemistry, 2016, 81(6): 2261-2279.

DOI：10.1021/acs.joc.5b02375

Although judicious use of chemical modifications has contributed to the success of nucleic acid therapeutics, poor systemic stability remains a major hurdle. The introduction of functional groups around the phosphate backbone can enhance the nuclease resistance of oligonucleotides (ONs). Here, we report the synthesis of enantiomerically pure (R)- and (S)-5'-C-methyl (C5'-Me) substituted nucleosides and their incorporation into ONs. These modifications generally resulted in a decrease in thermal stability of oligonucleotide (ON) duplexes in a manner dependent on the stereoconfiguration at C5' with greater destabilization characteristic of (R)-epimers. Enhanced stability against snake venom phosphodiesterase resulted from modification of the 3' end of an ON with either (R)- or (S)-C5'-Me nucleotides. The (S)-isomers with different 2'-substituents provided greater resistance against 3'-exonucleases than the corresponding (R)-isomers. Crystal structure analyses of RNA octamers with (R)- or (S)-5'-C-methyl-2'-deoxy-2'-fluorouridine [(R)- or (S)-C5'-Me-2'-FU, respectively] revealed that the stereochemical orientation of the C5'-Me and the steric effects that emanate from the alkyl substitution are the dominant determinants of thermal stability and are likely molecular origins of resistance against nucleases. X-ray and NMR structural analyses showed that the (S)-C5'-Me epimers are spatially and structurally more similar to their natural 5' nonmethylated counterparts than the corresponding (R)epimers.

• 出版日期2016-3-18