Synthesis of antisense oligonucleotides carrying modified 2-5A molecules at their 5'-termini and their properties.
The synthesis of 8-methyladenosine-substituted 2-5A tetramers with hydroxyalkyl groups at the 5'-phosphates and the corresponding 2-5A-antisense chimeras is described. These oligonucleotides were synthesized by the phosphoramidite method with a DNA/RNA synthesizer. These 2-5A tetramers with hydroxyethyl and hydroxybutyl groups at their 5'-phosphates were more resistant to hydrolysis by alkaline phosphatase than those without the hydroxyalkyl groups. Incorporation of the hydroxyethyl group into the 2-5A tetramer and 2-5A-antisense chimera slightly reduced the abilities of their analogues to activate recombinant human RNase L, but the abilities of the 2-5A tetramer and the 2-5A-antisense chimera both with the hydroxyethyl group and 8-methyladenosine returned to 80 and 50% relative to those of the oligonucleotides without the hydroxyethyl group and 8-methyladenosine, respectively. Furthermore, the enzyme activated by 8-methyladenosine-substituted 2-5A-antisense chimera with the hydroxyethyl group cleaved the complementary RNA as efficiently as that activated by 2-5A-antisense chimera without the hydroxyethyl group and 8-methyladenosine. Thus, the 2-5A-antisense chimera carrying the hydroxyethyl group and 8-methyladenosine will be a candidate for a novel antisense molecule.
Ueno
Yoshihito
Y
Department of Biomolecular Science, Faculty of Engineering, Gifu University, Yanagido, Gifu 501-1193, Japan.
Kato
Yoichiro
Y
Okatani
Shusaku
S
Ishida
Norihisa
N
Nakanishi
Masayuki
M
Kitade
Yukio
Y
D000227:Adenine Nucleotides / Q000138:chemical synthesis*; D004722:Endoribonucleases / Q000737:chemistry; D004789:Enzyme Activation; D006801:Humans; D009691:Nucleic Acid Denaturation; D016376:Oligonucleotides, Antisense / Q000138:chemical synthesis* / Q000378:metabolism; D009843:Oligoribonucleotides / Q000138:chemical synthesis*; D018075:RNA, Complementary / Q000737:chemistry; D011994:Recombinant Proteins / Q000138:chemical synthesis
D000227:Adenine Nucleotides; D016376:Oligonucleotides, Antisense; D009843:Oligoribonucleotides; D018075:RNA, Complementary; D011994:Recombinant Proteins; C023505:2',5'-oligoadenylate; D004722:Endoribonucleases; C031956:2-5A-dependent ribonuclease