Chemical synthesis and biological characterization of phosphorothioate analogs of 2', 5'-3'-deoxyadenylate trimer.
In continued studies to elucidate the requirements for binding to and activation of the 2',5'-oligoadenylate (2-5A) dependent endoribonuclease (RNase L), four 2-5A trimer analogs were examined to evaluate the effect of chirality of phosphorothioate substitution on biological activity. The chemical syntheses and purification of the four isomers of P-thio-3'-deoxyadenylyl-(2'-5')-P-thio-3'- deoxyadenylyl-(2'-5')-3'-deoxyadenosine, by the phosphoramidite approach, is described. The isolated intermediates were characterized by elemental and spectral analyses. The fully deblocked compounds were characterized by 1H and 31P NMR and HPLC analyses. The 2',5'-(3'dA)3 cores with either Rp or Sp chirality in the 2',5'-internucleotide linkages will bind to but will not activate RNase L. This is in contrast to 2',5'-A3 core analogs with either RpRp or SpRp phosphorothioate substitution in the 2',5'-internucleotide linkages which can bind to and activate RNase L. There are also marked differences in the ability of the 2',5'-A3 analogs to activate RNase L following introduction of the 5'-monophosphate. For example, the 5'monophosphates of 2',5'-(3'dA)3-RpRp and 2',5'-(3'dA)3-SpRp can bind to and activate RNase L, whereas the 5'-monophosphates of 2',5'-(3'dA)3-RpSp and 2',5'-(3'dA)3-SpSp can bind to but can not activate RNase L.
Sobol
R W
RW
Department of Biochemistry, Temple University School of Medicine, Philadelphia, PA 19140.
Charubala
R
R
Pfleiderer
W
W
Suhadolnik
R J
RJ
D001667:Binding, Competitive; D002851:Chromatography, High Pressure Liquid; D003838:Deoxyadenine Nucleotides / Q000138:chemical synthesis* / Q000378:metabolism / Q000494:pharmacology; D004722:Endoribonucleases / Q000378:metabolism*; D004789:Enzyme Activation / Q000187:drug effects; D054303:HIV Reverse Transcriptase; D015497:HIV-1 / Q000187:drug effects / Q000201:enzymology / Q000502:physiology; D009682:Magnetic Resonance Spectroscopy; D015394:Molecular Structure; D010710:Phosphates / Q000378:metabolism / Q000494:pharmacology; D010766:Phosphorylation; D018894:Reverse Transcriptase Inhibitors; D013237:Stereoisomerism; D013329:Structure-Activity Relationship; D014779:Virus Replication / Q000187:drug effects
D003838:Deoxyadenine Nucleotides; D010710:Phosphates; D018894:Reverse Transcriptase Inhibitors; D054303:HIV Reverse Transcriptase; D004722:Endoribonucleases; C031956:2-5A-dependent ribonuclease