Inhibition of hepatitis B virus DNA polymerase by 3′‐fluorothymidine triphosphate and other modified nucleoside triphosphate analogs

Meisel, Helga; Reimer, K.; Janta-Lipinski, M. Von; Bärwolff, D.; Matthes, E.

doi:10.1002/jmv.1890300211

Cited by 20 publications

(23 citation statements)

References 16 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…For estimation of the HBV DNA polymerase activity, virus particles were pelleted from HBV DNA positive human sera and the endogenous DNA polymerase was assayed as described previously (Meisel et al, 1990). The reaction mixture (49 ILl)contained 42mM Tris-hydrochloride (pH 7.5), 34mM MgCI2, 340mM KCI, 22 mM 2-mercaptoethanol, 0.4% Nonidet P40, 70 ILM of the fourth one as labelled triphosphate.…”

Section: Endogenous Hbv Dna Polymerasementioning

confidence: 99%

Inhibitory Effects of Acyclic Nucleoside Phosphonate Analogues on Hepatitis B Virus DNA Synthesis in HB611 Cells

Yokota¹,

Konno²,

Shigeta

et al. 1994

Antivir Chem Chemother

View full text Add to dashboard Cite

SummaryBy using an assay system based on a human hepatoblastoma cell line (HB611) that continuously synthesizes hepatitis B virus (HBV) DNA, 56 acyclic nucleoside phosphonate analogues were examined for their inhibitory effects on HBV DNA synthesis. The following compounds were found to inhibit HBV DNA synthesis at concentrations that were significantly lower than their minimum cytotoxic concentrations; 9-(2-phosphonylmethoxyethyl)adenine (PMEA), 9-(2-phosphonylmethoxyethyl) guanine(PMEG), 9-(2-phosphonylmethoxyethyl) guanine ethyl ester (PMEGEE), 9 -(2 -phosphonylmethoxyethyQ -1 -deazaadenine (PMEC 1A), 9-(2-phosphonylmethoxyethyl)-2,6-diaminopurine (PMEDAP), (S)-9-(3-hydroxy-2-phosphonylmethoxypropyl)adenine (HPMPA), 9-(3-isopropoxy-2-phosphonylmethoxypropyl)adenine (IPPMPA), 9-(RS)-(2-phosphonylmethoxypropyl)adenine (PMPA) and 9-(3-hydroxy-2-phosphonylmethoxypropyl)-2, 6-diaminopurine (HPMPDAP). The most selective compounds (with indexes greater than 100) were PMEDAP, PMEA, IPPMPA, and PMPA. Acyclic pyrimidine nucleoside phosphonate analogues did not prove markedly selective as anti-HBV agents. Diphosphoryl derivatives of some acyclic purine nucleoside phosphonates (i.e. PMEA, PMEDAP, HPMPA) were prepared. They proved inhibitory to HBV DNA polymerase but not cellular DNA polymerase ct.

show abstract

Section: Endogenous Hbv Dna Polymerasementioning

confidence: 99%

Inhibitory Effects of Acyclic Nucleoside Phosphonate Analogues on Hepatitis B Virus DNA Synthesis in HB611 Cells

Yokota¹,

Konno²,

Shigeta

et al. 1994

Antivir Chem Chemother

View full text Add to dashboard Cite

show abstract

“…The DNA polymerase a, the de novo DNA-synthesizing enzyme (1), is almost insensitive toward the triphosphate of FddThd. Recently, it was found that the triphosphates of the other nucleoside analogs studied here, FddMeCyt and ddeThd, were equally effective and selective inhibitors of HBV DNA polymerase (14).…”

Section: Discussionmentioning

confidence: 88%

“…While a 50o inhibition of DNA polymerase p was measured at a concentration of 2.2 ,uM triphosphate of FddThd, the inhibitory concentration of the triphosphate in the DNA polyrnerase a assay was >200 ,uM. In contrast, reverse trans-criptase from human immunodeficiency virus is inhibited at a concentration of as low as 0.05 ,uM (12) -and the-polymerase from HBV is inhibited at 0.15 p.M (14). This means that the cellular DNA polymerase ,, which is primarily involved in DNA repair (1), is 45-fold less sensitive to inhibition by the compound than the human immunodeficiency virus reverse transcriptase is and 15-fold less sensitive than the HBV DNA polymerase is.…”

Section: Discussionmentioning

confidence: 98%

Potent inhibition of hepatitis B virus production in vitro by modified pyrimidine nucleosides

Matthes

Langen

Janta-Lipinski

et al. 1990

Antimicrob Agents Chemother

View full text Add to dashboard Cite

2',3'-Dideoxy-3'-fluorothymidine (FddThd), 2',3'-didehydro-2',3'-dideoxythymidine (ddeThd), and 3'-fluoro-5-methyl-deoxycytidine (FddMeCyt) are, in their triphosphate forms, selective inhibitors of human immunodeficiency virus type 1 reverse transcriptase. We report that 0.3 ,uM FddThd, FddMeCyt, or ddeThd as well as 3'-chloro-5-methyl-deoxycytidine (ClddMeCyt) or 3'-amino-5-methyl-deoxycytidine (AddMeCyt) almost completely blocked production of hepatitis B virus (HBV) particles by HBV DNA-transfected cell line 2.2.15 in vitro. Only at an at least 10-fold-higher concentration was a cytotoxic effect observed. These results indicate that FddThd, FddMeCyt, ClddMeCyt, AddMeCyt, and ddeThd are potent anti-HBV agents in vitro.Hepatitis B is a disease with continuously increasing occurrence and is caused by the hepatitis B virus (HBV) (11). Vaccination against hepatitis B is one way of effectively preventing HBV infection (2). Two lines of treatment of hepatitis B infection have been followed: (i) treatment with cytokines (8) to substitute the impaired production of peripheral blood mononuclear cells (7, 32) and (ii) treatment with antiviral agents, e.g., adenine arabinoside (18,21,29). The rationale for a chemotherapeutic treatment for hepatitis B is the inhibition of the viral DNA polymerase (4).Recently, we found that the triphosphate of 2',3'-dideoxy-3'-fluorothymidine (FddThd) is a potent inhibitor of reverse transcriptase of human immunodeficiency virus, a weak inhibitor of cellular DNA polymerase ,, and not effective at all against DNA polymerase a (12). In addition, we established that FddThd is well phosphorylated to the 5'-triphosphate in relevant cell lines (13). Moreover, it was found that the triphosphate of FddThd and the triphosphates of 2',3'-didehydro-2',3'-dideoxythymidine (ddeThd) and 3'-fluoro-5-methyl-deoxycytidine (FddMeCyt) FddThd, FddMeCyt, ClddMeCyt, AddMeCyt, and ddeThd were synthesized as described previously (6, 9, 30; E. Matthes, C. Lehmann, D. Scholz, M. von Janta-Lipinski, K. Gaertner, P. Langen, and H. A. Rosenthal, European Patent 0254268A2, 1987).In vitro assay for antiviral activity. The human hepatoblastoma cell line HepG2 was transfected with pDolTHBV-1, a vector which contains HBV (27). The clonal line of cells was designated 2.2.15 and was found to secrete both hepatitis B surface antigen (HBsAg) and HBV DNA (27). The 2.2.15 cells were kept in RPMI 1640 medium supplemented with 2 mM glutamine and 10% (vol/vol) fetal bovine serum. The cultures were incubated at 37°C in 5% CO2 in air.Cells were planted at a density of 5 x 105/ml in plastic flasks. One hour later the compounds were added and incubation was continued for 4 days; the cultures reached confluence on day 5. On day 2 fresh serum was added to the culture medium and the respective compound concentration was renewed.Cell growth was determined by two methods. First, after the 4-day incubation period, the cultures were incubated for an additional 24 h with 3 pXCi of [3H]dAdo per ml. In order to avoid direct interference of th...

show abstract

“…Meanwhile, however, 3'-azido-2',3'-dideoxythymidine has been shown to be unable to suppress duck HBV (DHBV) replication in vitro and in vivo (9,26) or to be effective against hepatitis B infections of AIDS patients coinfected with HBV (4). A series of compounds, such as 2',3'-dideoxy-3'-fluorothymidine 5'-triphosphate and some closely related 3'-fluoro-modified dTTP analogs, 2',3'-didehydro-2',3'-dideoxythymidine 5'-triphosphate, ddTTP, ddCTP, ddGTP, and 5-substituted dUTP derivatives, were likewise described as efficient inhibitors of human immunodeficiency virus RT (1,8,18,19,24), exhibiting promising effects on HBV DNA polymerase (20,25) or DHBV DNA polymerase (14,25). Of these agents, only the 2',3'-dideoxynucleosides have been examined for their antiviral efficiency in two different HBV-producing cell systems (3,13,26) or in DHBV-infected animals (11,13), with varying success.…”

mentioning

confidence: 99%

“…The sugar-modified nucleosides and their corresponding 5'-triphosphates were synthesized and purified by wellestablished procedures (10,12) (20). The reaction mixture (49 ,ul) contained 42 mM Tris-HCl (pH 7.5), 34 mM MgCl2, 340 mM KCl, 22 mM 2-mercaptoethanol, 0.4% Nonidet P-40, 70 ,uM concentrations each of three of the deoxynucleoside triphosphates and, on the basis of the inhibitor examined, a 0.7 ,uM concentration of the fourth one as a labeled triphosphate (this concentration varied between 0.22 and 1.18 ,M in kinetic studies) (64 kdpm/pmol).…”

mentioning

confidence: 99%

Comparative inhibition of hepatitis B virus DNA polymerase and cellular DNA polymerases by triphosphates of sugar-modified 5-methyldeoxycytidines and of other nucleoside analogs

Matthes

Reimer

Janta-Lipinski

et al. 1991

Antimicrob Agents Chemother

View full text Add to dashboard Cite

Of a series of 14 nucleoside 5'-triphosphates, those of 2',3'-dideoxy-3'-fluoro-5-methylcytidine, 3'-azido-2',3'-dideoxy-5-methylcytidine, 2',3'-dideoxy-3'-fluoroguanosine, 2',3'-didehydro-2',3'-dideoxy-5-methylcytidine, 2',3'-dideoxy-3'-fluoro-5-ethylcytidine, and 2',3'-dideoxy-3'-fluoroadenosine emerged as the most potent inhibitors of hepatitis B virus DNA polymerase (50% inhibitory dose, 0.03 to 0.35 microM). In contrast, cellular DNA polymerases proved to be resistant to (alpha) or partially affected by (beta) these analogs. These compounds are among the most effective and selective inhibitors of endogenous hepatitis B virus DNA polymerase recognized to date.

show abstract

Inhibition of hepatitis B virus DNA polymerase by 3′‐fluorothymidine triphosphate and other modified nucleoside triphosphate analogs

Cited by 20 publications

References 16 publications

Inhibitory Effects of Acyclic Nucleoside Phosphonate Analogues on Hepatitis B Virus DNA Synthesis in HB611 Cells

Inhibitory Effects of Acyclic Nucleoside Phosphonate Analogues on Hepatitis B Virus DNA Synthesis in HB611 Cells

Potent inhibition of hepatitis B virus production in vitro by modified pyrimidine nucleosides

Comparative inhibition of hepatitis B virus DNA polymerase and cellular DNA polymerases by triphosphates of sugar-modified 5-methyldeoxycytidines and of other nucleoside analogs

Contact Info

Product

Resources

About