Inhibition of Hepatitis B Virus Replication by Nucleoside Enantiomers of β-2′,3′-Dideoxypurine Analogues

Alaoui, A.M. El; Faraj, Abdesslem; Pierra, Claire; Boudou, V.; Johnson, R. Jeremy; Mathé, Christophe; Gosselin, Gilles; Korba, Brent E.; Imbach, Jean‐Louis; Schinazi, Raymond F.; Sommadossi, Jean Pierre

doi:10.1177/095632029600700508

Cited by 18 publications

(4 citation statements)

References 23 publications

(21 reference statements)

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“…Finally, the rate of ␤-L-ddA catabolism can explain its differential antiviral activity against HIV (EC 50 , Ͼ100 M) grown in PBMC (1) and HBV (EC 50 , 5 to 6 M) grown in HepG2 cells (4,8). In PBMC, the metabolites resulting from [ 3 H]␤-LddA catabolism accounted for over 90% of the intracellular radioactivity (Table 2), while in HepG2 cells these metabolites accounted for only 40% of the total radioactivity (Table 1).…”

Section: Coincubation Of [ 3 H]␤-l-dda and Mta In Hepatocytes In Cultmentioning

confidence: 99%

“…135, 1998). However, ␤-L-ddA per se has a limited anti-HBV activity (50% effective concentration [EC 50 ], 5 to 6 M) in HBV DNA-transfected human hepatoblastoma-derived HepG2 cells (2.2.15 cells) and no anti-HIV activity (EC 50 , Ͼ100 M) in peripheral blood mononuclear cells (PBMC) (1,4,8,10). Previous studies showed that ␤-L-ddA is phosphorylated by 2Ј-deoxycytidine kinase (EC 2.7.1.74) with a high K m of 220 M (6).…”

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confidence: 99%

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Intracellular Metabolism of β- l -2′,3′-Dideoxyadenosine: Relevance to Its Limited Antiviral Activity

Placidi

Cretton-Scott

Gosselin

et al. 2000

Antimicrob Agents Chemother

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.28).Recent findings have indicated that several ␤-L-nucleoside analogs exhibit high and potent antiviral activity against both human immunodeficiency virus (HIV) (12) and hepatitis B virus (HBV) (2, 3, 5, 9, 13) replication accompanied by low host cellular toxicity when compared to their respective natural ␤-D-counterparts (15). These findings prompted a search for novel ␤-L-nucleoside derivatives which could have synergistic activities and/or do not exhibit cross-resistance with currently available chemotherapeutic nucleoside analogs so that they could be used in combination therapy regimens. We have found that ␤-L-dideoxyadenosine (ddA)-5Ј-triphosphate (␤-LddATP) is a potent inhibitor of both HIV type 1 reverse transcriptase and woodchuck hepatitis virus DNA polymerase (A. Faraj, L. Placidi, C. Perigaud, E. Cretton-Scott, G. Gosselin, L. T. Martin, C. Pierra, R. F. Schinazi, J. L. Imbach, and J. P. Sommadossi, Prog. Abstr. 13th Int. Round Table Nucleosides Nucleotides Biol. Appl., abstr. 135, 1998). However, ␤-L-ddA per se has a limited anti-HBV activity (50% effective concentration [EC 50 ], 5 to 6 M) in HBV DNA-transfected human hepatoblastoma-derived HepG2 cells (2.2.15 cells) and no anti-HIV activity (EC 50 , Ͼ100 M) in peripheral blood mononuclear cells (PBMC) (1,4,8,10). Previous studies showed that ␤-L-ddA is phosphorylated by 2Ј-deoxycytidine kinase (EC 2.7.1.74) with a high K m of 220 M (6). In addition, ␤-L-ddA is not a substrate for the catabolic enzyme purine nucleoside phosphorylase (EC 2.4.2.1) (11). The purpose of the present study was to investigate the intracellular metabolism of ␤-LddA in HepG2 cells, PBMC, and primary cultured hepatocytes in order to understand its limited in vitro antiviral activity. MATERIALS AND METHODS ␤-L-ddA was synthesized as previously described (10). [2Ј,3Ј,8-3 H]␤-L-ddA (18.2 mCi/mmol) was obtained by tritium reduction of ␤-L-2Ј,3Ј-didehydro-2Ј,3Ј-dideoxyadenosine (Moravek Biochemical). [2Ј,3Ј,8-3 H]␤-L-ddA was prepared by heterogeneous catalytic exchange with tritium gas in the presence of palladium catalyst and was Ͼ96% pure as ascertained by the high-performance liquid chromatography (HPLC) method described below. The presence of the tritium in both the base and L-dideoxyribose allowed us to follow the metabolism of this molecule.Cell culture conditions and preparation of samples. HepG2 cells were grown in 225-cm 2 tissue culture flasks in minimal essential medium with nonessential amino acids supplemented with 10% heat-inactivated dialyzed fetal bovine serum (FBS), 1% sodium pyruvate, and 1% penicillin-streptomycin. The medium was changed every 3 days, and the cells were subcultured once a week. After detachment of the adherent monolayer with a 10-min exposure to 30 ml of trypsin-EDTA and three consecutive washes with medium, confluent HepG2 cells (2 ϫ 10 6 /ml) were resuspended in a final volume of 10 ml of medium per time period and exposed to 10 M [ 3 H]␤-L-ddA (1,000 dpm/pmol). The cells were maintained at 37°C under a 5% CO 2 atmosphere for specified ti...

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Section: Coincubation Of [ 3 H]␤-l-dda and Mta In Hepatocytes In Cultmentioning

confidence: 99%

mentioning

confidence: 99%

Intracellular Metabolism of β- l -2′,3′-Dideoxyadenosine: Relevance to Its Limited Antiviral Activity

Placidi

Cretton-Scott

Gosselin

et al. 2000

Antimicrob Agents Chemother

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“…Previous studies have demonstrated that β-L-ddA lacked potent anti-HIV and anti-HBV activity in PBMCs and in 2.2.15 cells (Bolon et al, 1996;El Alaoui et al, 1996). The poor antiviral activity of β-L-ddA was attributed to a rapid and extensive catabolism of the drug by 5′-methylthioadenosine phosphorylase (EC 2.4.2.28).…”

Section: Discussionmentioning

confidence: 99%

Antiviral Activity and Intracellular Metabolism of Bis(tButylSATE) Phosphotriester of β-L-2′,3'Dideoxyadenosine, a Potent Inhibitor of HIV and HBV Replication

Placidi

Faraj

Ag³

et al. 2001

Antivir Chem Chemother

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The beta-L-nucleoside analogue beta-L-2',3'-dideoxy adenosine (beta-L-ddA) has been shown to exhibit limited antiviral activities. This was attributed to its rapid catabolism through cleavage of the glycosidic bond and poor phosphorylation to the nucleotide beta-L-2',3'-dideoxyadenosine-5'-mono phosphate (beta-L-ddAMP) (Placidi et al., 2000). However, the nucleotide beta-L-2',3'-dideoxyadenosine-5'-triphosphate (beta-L-ddATP) inhibited the activity of both HIV-1 reverse transcriptase (RT) and viral DNA polymerase isolated from woodchuck hepatitis virus-infected serum (a model of hepatitis B) with an inhibitory concentration (IC50) of 2.0 microM without inhibiting human DNA polymerases alpha, beta, or gamma up to a concentration of 100 microM. These results suggested that prodrugs of beta-L-ddAMP may bypass the poor metabolic activation of beta-L-ddA and lead to more potent and selective antiviral activity. Therefore, the mononucleoside phosphotriester derivative of beta-L-ddAMP incorporating the S-pivaloyl-2-thioethyl (tButylSATE) groups, beta-L-ddAMP-bis(tButylSATE) was synthesized. Beta-L-ddAMP-bis(tButylSATE) inhibited HIV replication in human peripheral blood mononuclear cells (PBMCs) and HBV replication in 2.2.15 cells with effective concentrations (EC50s) of 2 and 80 nM, respectively. Intracellular metabolism of beta-L-ddAMP-bis(tButylSATE) demonstrated that beta-L-ddATP was the predominant intracellular metabolite in PBMC and liver cells. The intracellular half-life of beta-L-ddATP was 5.4 and 9.2 h in HepG2 and PBMCs, respectively. The intracellular concentrations of beta-L-ddATP were maintained above the EC50 for the inhibition of HIV RT and hepatitis B virus (HBV) for as long as 24 h after removal of the drug.

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“…Nucleoside analogues have been a point of interest for chemists and biochemists due to their role in nucleic acid biosynthesis and various other biologically significant processes such as viral replication and division of cells. 1 In the past few decades, a variety of artificial or modified nucleosides have been synthesized by chemists to incorporate them in the antisense oligonucleotides [2][3][4] or to screen their biological activities such as antiviral, 5,6 anti-HIV, 7,8 anticancer, 9,10 antimetabolites, 11,12 antisense properties, 13 and many more. [14][15][16] Conformationally restricted nucleoside analogues with reduced conformational flexibility constituted a major class of these modified nucleosides.…”

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confidence: 99%

Chemoenzymatic Synthesis of arabino-Configured Bicyclic Nucleosides

et al. 2023

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A convergent route for the synthesis of a new class of bicyclic nucleosides has been developed. The synthetic route to the corresponding arabino-configured uracil and thymine bicyclic nucleosides proceeds in 24 and 27% overall yields, respectively, starting from 1,2,5,6-di-O-isopropylidene-α-d-glucofuranose. This synthetic protocol includes some crucial steps such as Vorbrüggen base coupling and chemo-enzymatic regioselective acetylation of the primary hydroxyl group by using Lipozyme® TL IM where it was found that Lipozyme® TL IM could be recovered and reused for selective acetylation without losing its selectivity.

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Inhibition of Hepatitis B Virus Replication by Nucleoside Enantiomers of β-2′,3′-Dideoxypurine Analogues

Cited by 18 publications

References 23 publications

Intracellular Metabolism of β- l -2′,3′-Dideoxyadenosine: Relevance to Its Limited Antiviral Activity

Intracellular Metabolism of β- l -2′,3′-Dideoxyadenosine: Relevance to Its Limited Antiviral Activity

Antiviral Activity and Intracellular Metabolism of Bis(tButylSATE) Phosphotriester of β-L-2′,3'Dideoxyadenosine, a Potent Inhibitor of HIV and HBV Replication

Chemoenzymatic Synthesis of arabino-Configured Bicyclic Nucleosides

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