A cell culture assay for compounds which inhibit hepatitis B virus replication

Korba, Brent E.; Milman, Gregory

doi:10.1016/0166-3542(91)90068-3

Cited by 105 publications

(50 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Discovery of novel antiviral compounds against HBV is hampered by a lack of cell-based assays suitable for high-throughput screening of compound libraries. Currently, the antiviral activity of compounds against HBV is usually assayed in human hepatoma (HepG2)-derived HBV replicating cell lines, such as HepG2.2.15 (35), HepAD38 (36), and Hep-DES19 (28), which typically require more than 6 days of incubation with drugs due to the slow dynamics of HBV replication in these cell lines (29,37,38). Recently, we established an immortalized mouse hepatocyte (AML12)-derived stable cell line (AML12HBV10) that supported HBV pgRNA transcription and subsequent DNA replication in a tetracycline (Tet)-dependent manner (27).…”

Section: Resultsmentioning

confidence: 99%

Sulfamoylbenzamide Derivatives Inhibit the Assembly of Hepatitis B Virus Nucleocapsids

Campagna

Liu

Mao

et al. 2013

J Virol

162

182

View full text Add to dashboard Cite

show abstract

Section: Resultsmentioning

confidence: 99%

Sulfamoylbenzamide Derivatives Inhibit the Assembly of Hepatitis B Virus Nucleocapsids

Campagna

Liu

Mao

et al. 2013

J Virol

162

182

View full text Add to dashboard Cite

show abstract

“…HepG2.2.15 cells were treated with lamivudine using a modification of a previously described method (19,20). Briefly, 4 ϫ 10 4 cells were grown to approximately 80 to 100% confluence in 24 wells and exposed to various concentrations of lamivudine.…”

Section: Methodsmentioning

confidence: 99%

Genomic Analysis of Anti-Hepatitis B Virus (HBV) Activity by Small Interfering RNA and Lamivudine in Stable HBV-Producing Cells

Guo

Zhang³

et al. 2005

J Virol

113

View full text Add to dashboard Cite

Hepatitis B virus (HBV) causes acute and chronic hepatitis and hepatocellular carcinoma. Small interfering RNA (siRNA) and lamivudine have been shown to have anti-HBV effects through different mechanisms. However, assessment of the genome-wide effects of siRNA and lamivudine on HBV-producing cell lines has not been reported, which may provide a clue to interrogate the HBV-cell interaction and to evaluate the siRNA's side effect as a potential drug. In the present study, we designed seven siRNAs based on the conserved HBV sequences and tested their effects on the expression of HBV genes following sorting of siRNA-positive cells. Among these seven siRNAs, siRNA-1 and siRNA-7 were found to effectively suppress HBV gene expression. We further addressed the global gene expression changes in stable HBV-producing cells induced by siRNA-1 and siRNA-7 by use of human genome-wide oligonucleotide microarrays. Data from the gene expression profiling indicated that siRNA-1 and siRNA-7 altered the expression of 54 and 499 genes, respectively, in HepG2.2.15 cells, which revealed that different siRNAs had various patterns of gene expression profiles and suggested a complicated influence of siRNAs on host cells. We further observed that 18 of these genes were suppressed by both siRNA-1 and siRNA-7. Interestingly, seven of these genes were originally activated by HBV, which suggested that these seven genes might be involved in the HBV-host cell interaction. Finally, we have compared the effects of siRNA and lamivudine on HBV and host cells, which revealed that siRNA is more effective at inhibiting HBV expression at the mRNA and protein level in vitro, and the gene expression profile of HepG2.2.15 cells treated by lamivudine is totally different from that seen with siRNA.

show abstract

“…However, there is currently no evidence of either intracellular phosphorylation of these compounds (7,8,18) DNA polymerase (22). This pattern of inhibition is distinctly different from that induced by inhibition of HBV replication by ara-AMP in 2.2.15 cells (a well-characterized inhibitor of DNA polymerase with no documented activity against HBV RT) in which a substantial accumulation of negative-strand DNA is observed (10). The present data are consistent with an inhibition of the HBV RT-directed priming step prior to elongation of the negative (first) strand of HBV DNA.…”

Section: Vol 48 2004 Anti-hbv Activities Of Di-and Trinucleotides 2201mentioning

confidence: 95%

Phosphorothioate Di- and Trinucleotides as a Novel Class of Anti-Hepatitis B Virus Agents

Iyer¹,

Jin²,

Roland³

et al. 2004

Antimicrob Agents Chemother

View full text Add to dashboard Cite

Several nucleoside analogs are under clinical development for use against hepatitis B virus (HBV). Lamivudine (3TC), a nucleoside analog, and adefovir dipivoxil (ADV), an acyclonucleotide analog, are clinically approved. However, long-term treatment can induce viral resistance, and following the cessation of therapy, viral rebound is frequently observed. There continues to be a need for new antiviral agents with novel mechanisms of action. A library of more than 600 di-and trinucleotide compounds synthesized by parallel synthesis using a combinatorial strategy was screened for potential inhibitors of HBV replication using the chronically HBV-producing cell line 2. The discovery of safe and effective antiviral drugs continues to present considerable challenges. The rapid emergence of resistance to antiviral drugs is a major problem, and combinations of a limited repertoire of antiviral drugs often need to be employed as a therapeutic strategy. The major stumbling block in antiviral drug development is the limited structural, mechanistic, and functional information on many virus-specific molecular targets. Consequently, the design of target-specific compounds that interfere with viral life cycles is a difficult challenge. There continues to be a substantial unmet clinical need for antiviral drugs with different structures and unique mechanisms of action, other than those conferred by conventional nucleoside analogs (2).Given a lack of sufficient structural information on new targets, an approach that seems appropriate for antiviral drug discovery is the screening of structurally diverse compounds, generated using combinatorial approaches that would modulate biological pathways without regard to specific molecular targets. In theory, this approach would allow simultaneous functional validation of a target, as well as the discovery of a lead structure that modulates the function of the target. This strategy has been variously referred to as "diversity-oriented organic synthesis for therapeutic target validation" or "combinatorial target-guided ligand assembly" (6, 20).We describe here the application of this concept for the discovery of anti-hepatitis B virus (HBV) agents. In recent publications, we have reported methods of assembling various classes of di-and trinucleotide libraries (7,8,18). Through screening, as well as lead optimization work, and in conjunction with cell-based assays, we have discovered that certain molecules show very promising anti-HBV activity. Furthermore, some of these molecules display synergistic activity when used in combination with lamivudine (3TC) and adefovir dipivoxil (ADV), the two nucleoside analogs currently licensed for the treatment of chronic HBV infection. This work summarizes the in vitro analysis and preliminary toxicity evaluation of these compounds.(A preliminary report of this work was presented at the 14th International Conference for Antiviral Research, Seattle, Washington, 8 to 12 April 2001.) MATERIALS AND METHODSTest compounds. Nucleotide libraries were synthesized ...

show abstract

A cell culture assay for compounds which inhibit hepatitis B virus replication

Cited by 105 publications

References 17 publications

Sulfamoylbenzamide Derivatives Inhibit the Assembly of Hepatitis B Virus Nucleocapsids

Sulfamoylbenzamide Derivatives Inhibit the Assembly of Hepatitis B Virus Nucleocapsids

Genomic Analysis of Anti-Hepatitis B Virus (HBV) Activity by Small Interfering RNA and Lamivudine in Stable HBV-Producing Cells

Phosphorothioate Di- and Trinucleotides as a Novel Class of Anti-Hepatitis B Virus Agents

Contact Info

Product

Resources

About