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2′,5′-Oligoadenylate-peptide nucleic acids (2–5A-PNAs) activate RNase L

1999; Elsevier BV; Volume: 7; Issue: 3 Linguagem: Inglês

10.1016/s0968-0896(98)00258-2

1464-3391

Jeroen C. Verheijen, Gijsbert A. van der Marel, Jacques H. van Boom, Suzanne F Bayly, Mark R. Player, Paul F. Torrence,

Advanced biosensing and bioanalysis techniques

To potentiate the 2–5A (2′,5′-oligoadenylate)-antisense and peptide nucleic acid (PNA) approaches to regulation of gene expression, composite molecules were generated containing both 2–5A and PNA moieties. 2–5A-PNA adducts were synthesized using solid-phase techniques. Highly cross-linked polystyrene beads were functionalized with glycine tethered through a p-hydroxymethylbenzoic acid linker and the PNA domain of the chimeric oligonucleotide analogue was added by sequential elongation of the amino terminus with the monomethoxytrityl protected N-(2-aminoethyl)-N-(adenin-1-ylacetyl)glycinate. Transition to the 2–5A domain was accomplished by coupling of the PNA chain to dimethoxytrityl protected N-(2-hydroxyethyl)-N-(adenin-1-ylacetyl)glycinate. Finally, (2-cyanoethyl)-N,N-diisopropyl-4-O-(4,4-dimethoxytrityl)butylphosphoramidite and the corresponding (2-cyanoethyl)-N,N-diisopropylphosphoramidite of 5-O-(4,4′-dimethoxytrityl)-3-O-(tert-butyldimethylsilyl)-N6-benzoyladenosine were the synthons employed to add the 2 butanediol phosphate linkers and the four 2′,5′-linked riboadenylates. The 5′-phosphate moiety was introduced with 2-[[2-(4,4′-dimethoxytrityloxy)ethyl]sulfonyl]ethyl-(2-cyanoethyl)-N,N-diisopropylphosphoramidite. Deprotection with methanolic NH3 and tetraethylammonium fluoride afforded the desired products, 2–5A-pnaA4, 2–5A-pnaA8 and 2–5A-pnaA12. When evaluated for their ability to cause the degradation of two different RNA substrates by the 2–5A-dependent RNase L, these new 2–5A-PNA conjugates were found to be potent RNase L activators. The union of 2–5A and PNA presents fresh opportunities to explore the biological and therapeutic implications of these unique approaches to antisense.