Inhibitory Effect of the Polyanionic Drug Suramin on the in Vitro HIV DNA Integration Reaction

Carteau, Sandrine; Mouscadet, Jean François; Goulaouic, Hélène; Subra, Frédéric; Auclair, Christian

doi:10.1006/abbi.1993.1468

Cited by 48 publications

(47 citation statements)

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“…We found that several polyhydroxylated anthraquinones were active against both purified integrase and PICs. Two tyrphostins, in contrast, were active against purified integrase (compounds 21 and 22), but were not active against PICs. Three anthraquinones, quinalizarin (compound 10), purpurin (compound 11), and alizarin (compound 12), inhibited purified PICs with IC50 values similar to those for inhibition of purified integrase (Table 1).…”

Section: Methodsmentioning

confidence: 95%

See 1 more Smart Citation

Differential inhibition of HIV-1 preintegration complexes and purified integrase protein by small molecules.

Farnet

Wang²,

Lipford³

et al. 1996

Proc. Natl. Acad. Sci. U.S.A.

114

123

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To replicate, HIV-1 must integrate a cDNA copy of the viral RNA genome into a chromosome of the host. The integration system is a promising target for antiretroviral agents, but to date no clinically useful integration inhibitors have been identified. Previous screens for integrase inhibitors have assayed inhibition of reactions containing HIV-1 integrase purified from an Escherichia coli expression system. Here we compare action of inhibitors in vitro on purified integrase and on subviral preintegration complexes (PICs) isolated from lymphoid cells infected with HIV-1. We find that many inhibitors active against purified integrase are inactive against PICs. Using PIC assays as a primary screen, we have identified three new anthraquinone inhibitors active against PICs and also against purified integrase. We propose that PIC assays are the closest in vitro match to integration in vivo and, as such, are particularly appropriate for identifying promising integration inhibitors.Following binding of HIV-1 to a sensitive cell, the viral and cellular membranes fuse and the viral core particle is released into the cytoplasm. There the viral genomic RNA is reverse transcribed, yielding a double-stranded DNA copy of the viral RNA genome. Next, the complex of viral cDNA and proteins, the "preintegration complex" (PIC), migrates to the nucleus and covalently attaches the viral cDNA to a chromosome of the host. Integration completes the formation of a provirus, which contains all the information necessary to direct the synthesis of the viral RNAs and proteins required for the formation of new virions (for reviews, see refs. 1 and 2). Fig. 1A summarizes the DNA breaking and joining reactions involved in integration in vivo. The blunt ends of the linear product of reverse transcription are cleaved to remove two nucleotides from each 3' end. The recessed 3' ends are then joined to protruding 5' ends of breaks made in the target DNA to yield an integration intermediate (Fig. 1A, II). The points of joining of the two DNA strands are staggered by five base pairs. The DNA between the two points of joining then melts, yielding gaps at each junction between host and viral DNA.

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Section: Methodsmentioning

confidence: 95%

“…1). Inhibition was monitored by Aurintricarboxylic acid (3) vuerIeIUgeIin to) Tyrphostin A51 (21) (Flavone) quantitating the reduction in the yield of integration product in the presence of drug.…”

Section: Methodsmentioning

confidence: 99%

Differential inhibition of HIV-1 preintegration complexes and purified integrase protein by small molecules.

Farnet

Wang²,

Lipford³

et al. 1996

Proc. Natl. Acad. Sci. U.S.A.

114

123

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“…However, since of all the pPT molecules tested T30177 maintained its level of enzyme inhibitory activity (Table 4), it is unlikely that the mechanism of inhibition is totally based on a polyanion effect, as is seen for compounds such as DS5000 and suramin (8). It is unclear at this time whether the G octet structure with the two-base-long T/dG loops found in T30177 is of paramount importance for inhibition of viral integrase since the G octet sequence found in T30659 did not inhibit integrase activity while T30526 (tetrad disrupting mutant) was able to inhibit enzyme activity, albeit at a reduced level.…”

Section: Discussionmentioning

confidence: 99%

T30177, an oligonucleotide stabilized by an intramolecular guanosine octet, is a potent inhibitor of laboratory strains and clinical isolates of human immunodeficiency virus type 1

Ojwang

Buckheit

Pommier

et al. 1995

Antimicrob Agents Chemother

114

112

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T30177, an oligonucleotide composed of only deoxyguanosine and thymidine, is 17 nucleotides in length and contains single phosphorothioate internucleoside linkages at its 5 and 3 ends for stability. This oligonucleotide does not share significant primary sequence homology with or possess any complementary (antisense) sequence motifs to the human immunodeficiency virus type 1 (HIV-1) genome. T30177 inhibited replication of multiple laboratory strains of HIV-1 in human T-cell lines, peripheral blood lymphocytes, and macrophages. T30177 was also found to be capable of inhibiting multiple clinical isolates of HIV-1 and preventing the cytopathic effect of HIV-1 in primary CD4 ؉ T lymphocytes. In assays with human peripheral blood lymphocytes there was no observable toxicity associated with T30177 at the highest concentration tested (100 M), while the median inhibitory concentration was determined to be in the range of 0.1 to 1.0 M for the clinical isolates tested, resulting in a high therapeutic index for this drug. In temporal studies, the kinetics of addition of T30177 to infected cell cultures indicated that, like the known viral adsorption blocking agents dextran sulfate and Chicago sky blue, T30177 needed to be added to cells during or very soon after viral infection. However, analysis of nucleic acids extracted at 12 h postinfection from cells treated with T30177 at the time of virus infection established the presence of unintegrated viral cDNA, including circular proviral DNA, in the treated cells. In vitro analysis of viral enzymes revealed that T30177 was a potent inhibitor of HIV-1 integrase, reducing enzymatic activity by 50% at concentrations in the range of 0.050 to 0.09 M. T30177 was also able to inhibit viral reverse transcriptase activity; however, the 50% inhibitory value obtained was in the range of 1 to 10 M, depending on the template used in the enzymatic assay. No observable inhibition of viral protease was detected at the highest concentration of T30177 used (10 M). In experiments in which T30177 was removed from infected cell cultures at 4 days post-HIV-1 infection, total suppression of virus production was observed for more than 27 days. PCR analysis of DNA extracted from cells treated in this fashion was unable to detect the presence of viral DNA 11 days after removal of the drug from the infected cell cultures. The ability of T30177 to inhibit both laboratory and clinical isolates of HIV-1 and the experimental data which suggest that T30177 represents a novel class of integrase inhibitors indicate that this compound is a viable candidate for evaluation as a therapeutic agent against HIV-1 in humans.One relatively new approach used in the development of antiviral therapeutics for human immunodeficiency virus type 1 (HIV-1) is the use of oligonucleotides designed as antisense agents (24,26,31). While much effort is being spent on rationally designed oligonucleotides such as antisense agents, there have also been recent findings of multiple alternative mechanisms by which oligonucleotides can i...

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“…The newly synthesized BDKAs exhibit potent inhibitory activity against IN for both ST and 3′-P steps. The acid derivatives (6,8) are more potent than the corresponding esters (5,7), and the 1-p-F-benzyl substituted quinolinones (7,8) are more active than the unsubstituted counterparts (5,6). Derivative 8 is the most potent derivative with IC 50 values for strand transfer around 15 nM ( Figure 3 and Table 1).…”

Section: Evaluation Of Biological Activitiesmentioning

confidence: 99%

Novel Bifunctional Quinolonyl Diketo Acid Derivatives as HIV-1 Integrase Inhibitors: Design, Synthesis, Biological Activities, and Mechanism of Action

et al. 2006

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The virally encoded integrase protein is an essential enzyme in the life cycle of the HIV-1 virus and represents an attractive and validated target in the development of therapeutics against HIV infection. Drugs that selectively inhibit this enzyme, when used in combination with inhibitors of reverse transcriptase and protease, are believed to be highly effective in suppressing the viral replication. Among the HIV-1 integrase inhibitors, the β-diketo acids (DKAs) represent a major lead for anti-HIV-1drug development. In this study, novel bifunctional quinolonyl diketo acid derivatives were designed, synthesized and tested for their inhibitory ability against HIV-1 integrase. The compounds are potent inhibitors of integrase activity. Particularly, derivative 8 is a potent IN inhibitor for both steps of the reaction (3′-processing and strand transfer) and exhibits both high antiviral activity against HIV-1 infected cells and low cytotoxicity. Molecular modeling studies provide a plausible mechanism of action, which is consistent with ligand SARs and enzyme photo-crosslinking experiments.

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Inhibitory Effect of the Polyanionic Drug Suramin on the in Vitro HIV DNA Integration Reaction

Cited by 48 publications

References 0 publications

Differential inhibition of HIV-1 preintegration complexes and purified integrase protein by small molecules.

Differential inhibition of HIV-1 preintegration complexes and purified integrase protein by small molecules.

T30177, an oligonucleotide stabilized by an intramolecular guanosine octet, is a potent inhibitor of laboratory strains and clinical isolates of human immunodeficiency virus type 1

Novel Bifunctional Quinolonyl Diketo Acid Derivatives as HIV-1 Integrase Inhibitors: Design, Synthesis, Biological Activities, and Mechanism of Action

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