Inhibition of HIV-1 replication and activation of RNase L by phosphorothioate/phosphodiester 2',5'-oligoadenylate derivatives.
2',5'-Oligoadenylate (2-5A) derivatives have been designed to act distal to the human immunodeficiency virus-1 (HIV-1)-induced blockade in the 2-5A synthetase/RNase L antiviral pathway. Stereochemical modification of individual internucleotide linkages of the 2-5A molecule was accomplished by phosphoramidite and phosphotriester chemical syntheses. Phosphorothioate/phosphodiester trimer and tetramer 2-5A derivatives revealed differences in the stereodynamics of activation of RNase L and inhibition of HIV-1 replication. The first and second internucleotide linkages are critical for activation of recombinant, human RNase L; A(Rp)ApA, A(Sp)ApA and ApA(Rp)A are agonists (IC50 = 2 x 10(-7), 2 x 10(-6) and 8 x 10(-6) M); ApA(Sp)A is an antagonist. The second and third internucleotide linkages are crucial for activation of murine RNase L; ApA(Rp)A, ApA(Rp)ApA, and ApApA(Rp)A are agonists (IC50 = 5 x 10(-7) M); ApA(Sp)A, ApA(Sp)ApA, and ApApA(Sp)A are antagonists. Inhibition of HIV-1-induced syncytia formation by the phosphorothioate/phosphodiester derivatives is specific for derivatives with substitution at the 2',3'-terminus. ApA(Rp)A, ApA(Sp)A, ApApA(Rp)A, and ApApA(Sp)A are potent inhibitors of HIV-1-induced syncytia formation (80-, 10-, 40-, and 15-fold more inhibitory, respectively, than solvent control). HIV-1 infection results in enhanced uptake and accumulation of ApA(Rp)A and ApA(Sp)A (7- and 10-fold, respectively). These stereochemically modified 2-5A derivatives are taken up preferentially by HIV-1-infected cells and show promise in anti-HIV-1 chemotherapy.
Sobol
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RW
Department of Biochemistry, Temple University School of Medicine, Philadelphia, Pennsylvania 19140.
Henderson
E E
EE
Kon
N
N
Shao
J
J
Hitzges
P
P
Mordechai
E
E
Reichenbach
N L
NL
Charubala
R
R
Schirmeister
H
H
Pfleiderer
W
W
D000227:Adenine Nucleotides / Q000138:chemical synthesis* / Q000378:metabolism / Q000494:pharmacology*; D000818:Animals; D000998:Antiviral Agents / Q000138:chemical synthesis / Q000494:pharmacology*; D001483:Base Sequence; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D017931:DNA Primers; D004722:Endoribonucleases / Q000096:biosynthesis / Q000378:metabolism*; D004789:Enzyme Activation; D004926:Escherichia coli; D015726:Giant Cells / Q000187:drug effects; D015497:HIV-1 / Q000187:drug effects / Q000502:physiology*; D006801:Humans; D007202:Indicators and Reagents; D007700:Kinetics; D007739:L Cells; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D015183:Restriction Mapping; D013329:Structure-Activity Relationship; D014779:Virus Replication / Q000187:drug effects*
D000227:Adenine Nucleotides; D000998:Antiviral Agents; D017931:DNA Primers; D007202:Indicators and Reagents; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D004722:Endoribonucleases; C031956:2-5A-dependent ribonuclease