Free accessResearch articleFirst published online 1996-4
Properties and Anti-Hiv Activity of Hairpin Antisense Oligonucleotides Containing 2′-Methoxynucleosides with Base-Pairing in the Stem Region at the 3′-End
A new type of hairpin antisense oligodeoxyribo-nucleotide, containing 2′-methoxynucleosides with base-pairing in the stem region at the 3′-end, was tested for 3′-exonuclease resistance and anti-HIV activity. An increased resistance to nuclease degradation has been observed by incubation of the hairpin oligo-nucleotides with DNA polymerase and foetal bovine serum. Of particular interest is the hairpin antisense oligonucleotide containing 2′-methoxynucleosides with base-pairing in the stem region at the 3′-end, which has increased nuclease resistance and hybridizes effectively with a complementary RNA. Furthermore, these compounds were assayed for inhibition of virus replication in HIV-1 infected MT-4 cells. In the anti-HIV activity test, the hairpin oligonucleotide phosphorothioate derivatives showed higher anti-HIV activities compared to their linear counterparts.
AgrawalS.GoodchildJ.CiveiraM.P.ThorntonA.H.SarinP.S., and ZamecnikP.C. (1988) Oligodeoxynucleoside phosphoramidites and phosphorothioates as inhibitors of human immunodeficiency virus. Proc Natl Acad Sci USA85: 7079–7083.
2.
AgrawalS.TemsamaniJ., and TangJ.-Y. (1991) Pharmacokinetics, biodistribution, and stability of oligodeoxynucleotide phosphorothioate in mice. Proc Natl Acad Sci USA88: 7595–7599.
3.
AgrawalS. (1992) Antisense oligonucleotides as antiviral agents. Trends in Biotechnology10:152–158.
BennettC.F., and CrookeS.T. (1994) Regulation of endothelial cell adhesion molecule expression with antisense oligonucleotides. In Advances in Pharmacology. AugustT.AndersM.W.MuradF., and CoyleJ.T. (eds). San Diego: Academic Press, 28, pp. 1–43.
6.
CalabrettaB. (1991) Inhibition of protoconcogene expression by antisense oligodeoxynucleotides: Biological and therapeutic implications. Cancer Res51: 4505–4510.
CrookeS.T. (1995) Therapeutic Applications of Oligonucleotides. New York: Springer Verlag.
9.
GhoshM.K., and ChoenJ.S. (1992) Oligodeoxynucleotides as antisense inhibitions of gene expression. Proc Nucleic Acids Res42: 79–126.
10.
GoodchildJ.AgrawalS.CiveraM.P.SarinP.S.SunD., and ZamecnikP.C. (1988) Inhibition of human immunodeficiency virus replication by antisense oligodeoxynucleotides. Proc Natl Acad Sci USA85: 5507–5511.
11.
HattaT.KimS.-G.SuzukiS.TakaiK., and TakakuH. (1994) Anti-Human immunodeficiency virus activity and mechanisms of unmodified and modified antisense oligonucleotides. In Carbohydrate modifications in antisense research. SanghviY.S., and CookP.D. (eds) Washington: American Chemical Society, pp. 151–168.
12.
HélèneC., and ToulméJ.J. (1990) Specific regulation of gene expression by antisense, sense, and antigene nucleic acids. Biochem Biophys Acta1049: 99–125.
13.
IversenP. (1991) In vivo studies with phosphorothioate oligonucleotide: pharmacokinetic prologue. Anti-Cancer Drug Design6: 531–538.
14.
JaskulskiD.DeRielJ.K.MercerW.E.CalabretaB., and BasergaR. (1989) Inhibition of cellular proliferation by antisense oligodeoxynucleotides to PCNA cyclin. Science240: 1544–1546.
15.
KimS.-G.SuzukiY.NakashimaH.YamamotoN., and TakakuH. (1991) Phosphorothioate analogues of oligodeoxyribonucleotides: Synthesis and activity as inhibitors of replication of human immunodeficiency virus. Biochem Biophys Res Commun179:1614–1619.
16.
KimS.-G.NakashimaH.YamamotoN., and TakakuH. (1993) Anti-HIV activities and physicochemical properties of phosphorothioate analogues complementary to HIV sequences. Bioorg Med Chem Lett3, 1223–1228.
17.
KimS.-G.HattaT.TsukaharaS.NakashimaH.YamamotoN.ShojhiY., and TakakuH. (1995) Antiviral effect of phosphorothioate oligodeoxyribonucleotides complementary to human immunodeficiency virus. Bioorg Med Chem3: 49–54.
18.
LemaitreM.BayardB., and LebleuB. (1987) Specific antiviral activity of a poly(L-lysine)-conjugated oligonucleotide sequence complementary to vesicular somatitis virus N protein mRNA initiation site. Proc Natl Acad Sci USA84: 648–652.
19.
LeonettiJ.P.DegolsG.ClarencJ.P.MechtiN., and LebleuB. (1993) Cell delivery and mechanisms of action of antisense oligonucleotide. Prog Nucleic Acids Res44:143–166.
20.
LetiterJ.M.E.AgrawalS.PaleseP., and ZamecnikP.C. (1990) Inhibition of influenza virus replication by phosphorothioate oligodeoxynucleotides. Proc Natl Acad Sci USA87: 3430–3434.
21.
LewinB. (1990) Driving the cell cycle: M phase kinase, its patterns and substrates. Cell61: 743–752.
22.
MatsukuraM.ShinozukaK.ZonG.MitsuyaH.ReitzM.CohenJ.C., and BroderS. (1987) Phosphorothioate analogs of oligodeoxynucleotides: Inhibitors of replication and cytopathic effects of human immunodeficiency virus. Proc Natl Acad Sci USA84: 7706–7710.
23.
MillerP.S.AgrisC.H.AurelainL.BlakeK.R.MurakamiA.ReddyM.P.SpitS.A. and Ts'oP.O.P. (1985) Control of ribonucleic acid function by oligonucleoside methyl-phosphonates. Biochemie67: 769–776.
24.
MurrayJ.A.H. (ed) (1992) Antisense RNA and DNA. New York: John Wiley & Sons.
25.
NakashimaH.ShojiY.KimS.-G.ShimadaJ.MizushimaY.ItoM.YamamotoN., and TakakuH. (1994) Anti-human immunodeficiency virus type I activity of phosphorothioate analogs of oligodeoxynucleotides: Penetration and Localization of oligonucleotides in HIV-1-infected MOLT-4 cells. Nucleic Acids Res22: 5004–5010.
26.
PauwelsR.BalzariniJ.BabaM.SnoeckR.ScholsD.HerdewijinP.DesmyterJ., and De ClercqE. (1988) Rapid and automated tetrazolium-based colorimetric assay for the detection of anti-HIV compounds. J Virol Methods20: 309–321.
27.
PrakashG., and KoolE.T. (1991) Molecular recognition by circular oligonucleotides. Strong binding of single-stranded DNA and RNA. J Chem Soc Chem Commun1161–1163.
28.
SmithC.C.AurelianL.ReddyM.P.MillerP.S., and Ts'oP.O.P. (1986) Antiviral effect of an oligo-(nucleoside methyl-phosphonate) complementary to the splice junction of herpes simplex virus type 1 immediate early pre-mRNAs 4 and 5. Proc Natl Acad Sci USA83: 2787–2791.
29.
ShimizuM.KonishiA.ShimadaY.InoueH., and OhtsukaE. (1992) Oligo(2′-O-methyl) ribonucleotides effective probes for duplex DNA. FEBS302:155–158.
30.
SteinC.A., and CohenJ.S. (1988) Oligodeoxynucleotides as inhibitions of gene expression. Cancer Res48: 2659–2668.
31.
TangJ.Y.TemsamaniJ., and AgrawalS. (1993) Self-stabilized antisense oligodeoxynucleotide phosphorothioate: Properties and anti-HIV activity. Nucleic Acids Res21: 2729–2735.
32.
ToulméJ.J., and HélèneC. (1988) Antimessenger oligodeoxynucleotides: an alternative to antisense RNA for artificial regulation of gene expression. Gene72: 51–58.
33.
TravaliS.ReissK.FerberA.PetraliaS.MercerW.E.CalabretaB., and BasergaR. (1991) Constitutively expressed c-myb abrogates the requirement for insulin-like growth factor 1 in 3T3 fibroblasts. Mol Cell Biol11: 731–736.
34.
ZamecnikP.C., and StephensonM.L. (1978) Inhibition of Rous sarcoma virus replication and cell transformation by a specific oligodeoxynucleotide. Proc Natl Acad Sci USA75: 280–284.
35.
ZamecnikP.C.GoodchildJ.TaguchiY., and SarinP.S. (1986) Inhibition replication and expression of human T-cell synthetic oligonucleotides complementary to viral RNA. Proc Natl Acad Sci USA83: 4143–4146.
36.
ZerialA.ThougNT., and HélèneC (1987) Selective inhibition of the cytopathic effect of type A influenza viruses by oligodeoxynucleotides covalently linked to an intercalating agent. Nucleic Acids Res15: 9909–9919.
37.
WickstromE. (ed) (1991) Prospects for Antisense Nucleic Acid Therapy for Cancer and AIDS. New York: John Wiley & Sons.
