Inhibitory effect of 9-(2-phosphonylmethoxyethyl)adenine (PMEA) on human and duck hepatitis B virus infection

Heijtink, R. A.; Wilde, Gemma; Kruining, J.; Berk, Luuk; Balzarini, Jan; Clercq, Erik De; Holý, Antonı́n; Schalm, Solko W.

doi:10.1016/0166-3542(93)90050-s

Cited by 122 publications

(86 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Similarly, it was reported that hepatitis B surface antigen production was not inhibited in HepG2 2.2.15 human hepatoma cell line during adefovir treatment, although similar experiments using DHBV-infected duck hepatocytes indicated a drug concentration-dependent inhibition of duck hepatitis B surface antigen production. 15 The lack of an inhibitory effect of adefovir on woodchuck hepatitis B surface antigen production was not surprising, because our data showed that adefovir treatment did not lead to reduction of the cccDNA, which is also the template for the synthesis of the preS and S RNAs. In fact, the persistence of cccDNA in the nucleus of infected hepatocytes implies that viral RNA levels may also remain constant, providing that our woodchuck primary hepatocyte cultures stayed highly differentiated.…”

Section: Discussionmentioning

confidence: 92%

“…The antiviral activity of PMEA against hepadnaviruses has been evaluated previously in human hepatoma cell lines and in the duck HBV (DHBV) model, where it appeared to be a potent inhibitor of hepadnavirus polymerases. 15,16 Furthermore, in recent phase II clinical trials, adefovir dipivoxyl, the oral pro-drug of PMEA, displayed a median 4.1-log decrease in plasma HBV-DNA levels. 17 Here we show that treatment of primary woodchuck hepatocytes with adefovir for up to 24 days inhibits WHV-DNA synthesis and secretion very efficiently.…”

mentioning

confidence: 99%

See 1 more Smart Citation

Increased hepatocyte turnover and inhibition of woodchuck hepatitis B virus replication by adefovirIn vitro do not lead to reduction of the closed circular DNA

et al. 2000

View full text Add to dashboard Cite

The aim of this study was to evaluate the inhibitory effect of the nucleotide analogue adefovir on woodchuck hepatitis B virus (WHV) replication and, in particular, to determine whether the pool of covalently closed circular DNA (cccDNA) could be reduced by adefovir treatment in primary cultures of woodchuck hepatocytes isolated from a chronic carrier. Strong reduction of WHV-DNA synthesis (90%) and secretion (up to 98%) was observed with all 3 doses of adefovir used (1, 10, and 100 mol/L), whereas in the absence of the drug, high amounts of viral particles were continuously secreted in the culture medium until the end of the study (27 days). Secretion of envelope proteins and viral RNA levels remained constant both in the adefovirtreated and -untreated cultures for the entire course of the study. Intracellular core protein levels declined by approximately 50% in all the cultures, independent of adefovir treatment. There was no indication of cccDNA loss in the adefovir-treated hepatocyte cultures even when cell turnover was induced for 14 days by the addition of epidermal growth factor (EGF) to the culture medium. Our data show that adefovir has a very strong inhibitory effect on WHV-DNA synthesis in chronically infected primary hepatocyte cultures and indicate that cccDNA is a very stable molecule that appears to be efficiently transmitted to the dividing hepatocytes. (HEPATOLOGY 2000;32:139-146.)Persistent infection with hepatitis B virus (HBV) continues to be a major health problem as it is associated with various degrees of liver damage, cirrhosis, and hepatocellular carcinoma. Despite the existence of an effective vaccine, more than 350 million people are infected worldwide. 1,2 Current treatment regimens primarily rely on interferon alfa therapy. However, interferon treatment is associated with a variety of side effects and has been successful in only approximately one third of the patients, being very poorly effective in individuals with perinatally acquired HBV infection. In recent years, a variety of new anti-HBV agents have been developed, and these are currently evaluated in clinical trials. 3,4 Because HBV replication proceeds through reverse transcription, treatment with inhibitors of HBV reverse transcriptase is a very attractive option.During hepadnavirus infection, the relaxed circular partially double stranded DNA genome is transported to the nucleus of the hepatocyte, where it is converted into a covalently closed circular molecule, named cccDNA, which serves as template for the transcription of all viral RNAs. 5 The synthesis of the new relaxed circular viral DNA genome is mediated by the viral P-protein, which has reverse transcriptase and DNA-dependent DNA polymerase activity. Synthesis of the viral DNA takes place in the cytoplasm of the hepatocytes within immature nucleocapsids. 6 To establish a sufficient pool of nuclear cccDNA, some of the newly synthesized viral DNAs need to be retransported to the nucleus of the infected hepatocyte. 7 Therefore, the replicative forms of viral DNA are a...

show abstract

Section: Discussionmentioning

confidence: 92%

mentioning

confidence: 99%

Increased hepatocyte turnover and inhibition of woodchuck hepatitis B virus replication by adefovirIn vitro do not lead to reduction of the closed circular DNA

et al. 2000

View full text Add to dashboard Cite

show abstract

“…A previous study (1) indicated that it has no clinically relevant activity against human immunodeficiency virus -type 1 (HIV), and this conclusion is included in the package insert. This selective activity against HBV is unique amongst the nucleoside/nucleotide analogues available for hepatitis B treatment (4)(5)(6)(7). Thus, several treatment guidelines, including the United States Public Health Service Guidelines, recommend entecavir for hepatitis B treatment in HIV-infected individuals who do not meet criteria for HIV treatment (8)(9)(10)(11).…”

Section: Introductionmentioning

confidence: 99%

The HBV Drug Entecavir — Effects on HIV-1 Replication and Resistance

et al. 2007

View full text Add to dashboard Cite

“…This colorimetric method allows for the quantitative detection of HBV DNA and can be partially automated. The use of HB611 cells for analysis of the anti-HBV compounds has been reported previously (4,8,14,17,18). However, we have developed a rapid and accurate colorimetric assay system for anti-HBV agents using HB611 cells, immunoaffinity purification, PCR, and hybrid-capture detection.…”

Section: Resultsmentioning

confidence: 99%

“…However, human hepatoma cell lines transfected with HBV DNA have also been developed (HB611 and HepG2 2.2.15 cells (11,13)) and used for the screening of antiviral drugs (3,4,8,14,17,18). In these studies, drug efficacy was determined by measuring HBV DNA levels using Southern blot analyses with 9)-labeled HBV DNA as the probe.…”

mentioning

confidence: 99%

Colorimetric Assay System for Screening Antiviral Compounds against Hepatitis B Virus

Sudo

Konno²,

Shigeta

et al. 1996

Microbiology and Immunology

View full text Add to dashboard Cite

A highly sensitive, rapid, and accurate assay system was developed for the in vitro evaluation of anti-hepatitis B virus (anti-HBV) agents. Chronic HBV-producing HB611 cells were used in combination with immunoaffinity purification, polymerase chain reaction (PCR), and hybrid capture detection. HB611 cells were incubated with putative anti-HBV agents for 7 days in 96-well microtiter plates. HBV was purified from HB611 cell culture media using immunoaffinity purification. The HBV DNA was extracted, amplified with PCR, and assayed using a hybrid capture colorimetric method. This assay provided quantitative detection of extracellular HBV DNA from 25 Al of cell culture media. Using the colorimetric method, we found that 50% effective concentration levels of several known anti-HBV agents (HPMPA, PMEDAP, PMEA and others) were similar to those reported in studies using Southern blot analysis. These results demonstrate that this new and easily automated colorimetric assay system can be used for the rapid and accurate assessment of anti-HBV compound selectivity.

show abstract

Inhibitory effect of 9-(2-phosphonylmethoxyethyl)adenine (PMEA) on human and duck hepatitis B virus infection

Cited by 122 publications

References 33 publications

Increased hepatocyte turnover and inhibition of woodchuck hepatitis B virus replication by adefovirIn vitro do not lead to reduction of the closed circular DNA

Increased hepatocyte turnover and inhibition of woodchuck hepatitis B virus replication by adefovirIn vitro do not lead to reduction of the closed circular DNA

The HBV Drug Entecavir — Effects on HIV-1 Replication and Resistance

Colorimetric Assay System for Screening Antiviral Compounds against Hepatitis B Virus

Contact Info

Product

Resources

About