Inhibition of Human Immunodeficiency Virus Type 1 Activity In Vitro by a New Self-Stabilized Oligonucleotide with Guanosine-Thymidine Quadruplex Motifs

Abstract

An oligonucleotide with a dimeric hairpin guanosine quadruplex (basket type structure) (dG3T4G3-s), containing phosphorothioate groups, was able to inhibit human immunodeficiency virus type 1 (HIV-1)-induced syncytium formation and virus production (as measured by p24 core antigen expression) in peripheral blood mononuclear cells. This oligonucleotide lacks primary sequence homology with the complementary (antisense) sequences to the HIV-1 genome. Furthermore, this oligonucleotide may have increased nuclease resistance. The activity of this oligonucleotide was increased when the phosphodiester backbone was replaced with a phosphorothioate backbone. In vivo results showed that dG3T4G3-s was capable of blocking the interaction between gp120 and CD4. We also found that dG3T4G3-s specifically inhibits the entry of T-cell line-tropic HIV-1 into cells. This compound is a viable candidate for evaluation as a therapeutic agent against HIV-1 in humans.

Inhibition of HIV-1 Replication by Oligonucleotide Analogues Directed to the Packaging Signal and Trans-Activating Response Region

“…HIV has been a frequent target for both RNAdegrading and steric-block approaches (Arzumanov et al, 2001b;Boulme et al, 1998;Darfeuille et al, 2002a;Darfeuille et al, 2004;Hiratou et al, 2001;Ittig et al, 2004;Kaushik et al, 2002;Park et al, 2000;Sei et al, 2000;Suzuki et al, 2002;Yamaguchi et al, 1997). For antisense technologies to become drug candidates, sequence specificity is essential.…”

Newly Designed Six-Membered Azasugar Nucleotide-Containing Phosphorothioate Oligonucleotides as Potent Human Immunodeficiency Virus Type 1 Inhibitors

“…a CC 50 , concentration required to inhibit cell survival 50%. Cells were incubated for 4 days in the presence of DBM ONs at different concentrations and then tested for their viability by MTT assay.…”

“…Cells in 96-well plates (2 ϫ 10 4 cells/well) were incubated with serially diluted DBM ONs for 4 days, and the number of viable cells was quantified via MTT assays. Cytotoxicity was assessed according to the concentration of ONs required to reduce cell viability by 50% (CC 50 ).…”

“…The mechanisms suggested have included adsorption blocking (9,16,47,50,51,53) and inhibition of HIV-1-specific enzymes, such as reverse transcriptase (29)(30)(31) or integrase (21-23, 36, 42). However, most studies of the antiviral mechanisms of PAS ONs, as well as recent studies with small interfering RNA (siRNA) against HIV-1 (3,5,7,35,38,39,49), have been conducted by transfection or viral vector-mediated delivery (4,17,27,33,34), rather than simple treatment of the infected culture.…”

Structure-based drug design: From nucleic acid to membrane protein targets

“…A notable example is the previously mentioned AS1411 (Ireson et al , 2006), which is described in detail by Bates et al in this issue. Quadruplexes also have been shown to have antiviral activity and have been demonstrated to be effective against HIV-1 in vivo (Bishop et al , 1996; Suzuki et al , 2002). Mainly due to their ability to recognize both nucleic acids and proteins with a high degree of specificity and because of their stability and nuclease resistance quadruplexes are rapidly becoming attractive targets for development of novel therapeutics and preclinical studies are underway for several other quadruplex-based therapeutics (Cogoi et al , 2006; Qi et al , 2006).…”