Phosphorylation, anti-HIV activity and cytotoxicity of 3′-fluorothymidine

Matthes, E.; Lehmann, Christian; Scholz, D.; Rosenthal, H. A.; Langen, P.

doi:10.1016/s0006-291x(88)81170-7

Cited by 83 publications

(35 citation statements)

References 10 publications

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“…FLT was phosphorylated better than AZT in all three cell types, despite the lower concentration of FLT used, which is presumably the basis for the higher antiviral activity and higher toxicity of FLT compared to AZT (Balzarini et al, 1988;Hartmann et al, 1988;Matthes et al, 1988;Bazin et al, 1989;Koshida et al, 1989a).…”

Section: Discussionmentioning

confidence: 91%

“…Both compounds were ph()spho~alled more efficiently in GEM cells than H9 cells, a fac::l Ylh~111 might be linked to the greater toxicity of the compounds in GEM cells (Balzarini et al, 1988;Matthes et al, 1988;Gox and Harmenberg, 1990). FLT was phosphorylated better than AZT in all three cell types, despite the lower concentration of FLT used, which is presumably the basis for the higher antiviral activity and higher toxicity of FLT compared to AZT (Balzarini et al, 1988;Hartmann et al, 1988;Matthes et al, 1988;Bazin et al, 1989;Koshida et al, 1989a).…”

Section: Discussionmentioning

confidence: 97%

“…The most studied of these, 3'-azido-3'-deoxythymidine (AZT), is used clinically to treat AIDS patients, and other dideoxynucleosides are currently undergoing clinical trials, including 3'-fluoro-3'-deoxythymidine (FLT) one of the most active dideoxynucleosides described to date (Hartmann et al, 1988;Matthes et al, 1988;Bazin et al, 1989;Koshida et a/., 1989a). These compounds are phosphorylated intracellularly (by the thymidine salvage pathway in the case of AZT and FLT) and, as the triphosphate, inhibit the HIV reverse transcriptase (RT) by chain termination, since they lack the 3'-hydroxyl necessary for further elongation (Furman et al, 1986;Matthes et al, 1988).…”

Section: Introductionmentioning

confidence: 99%

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Metabolism of 3′-Azido-3′-Deoxythymidine and 3′-Fluoro-3′-Deoxythymidine in Combination in Human Immunodeficiency Virus Infected Lymphoblastoid Cells

Cox

1992

Antivir Chem Chemother

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SummaryA combination of 3'-azido-3'-deoxythymidine (AZT) with 3'-fluoro-3'-deoxythymidine (FLT) has been shown previously to give synergistic inhibition of human immunodeficiency virus replication and greatly reduced cytotoxicity in vitro. The phosphorylation of the compounds, and their effect upon the natural deoxynucleoside triphosphate pools, were compared in CEM, H9, and HIV-infected H9/ymphoblastoid cells, both for the compounds when used alone and when combined together.Higher levels of FLT triphosphate than AZT triphosphate, and higher levels of AZT monophosphate than FLT monosphosphate, were formed in all cell types. Both compounds were phosphorylated most efficiently in CEM cells, whereas they were least efficiently phosphorylated in infected H9 cells.Owing to competition, the phosphorylation of both analogues was reduced when used in combination, compared to the phosphorylation of the separate compounds. The phosphorylation of the separate compounds was therefore at a maximum and was not increased by combining the compounds. The two compounds competed equally with each other for phosphorylation when used at a ratio of AZT: FLT of 5:1.Both analogues severely reduced the deoxynucleoside triphosphate pools in uninfected and human immunodeficiency virus-infected H9 cells, but not in CEM cells. The effects of the two compounds were similar to those found when the compounds were combined, and thus H9 cells were shown to be much more sensitive to the effects of the analogues upon deoxynucleoside triphosphate pools than CEM cells were.Received

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Section: Discussionmentioning

confidence: 91%

Section: Discussionmentioning

confidence: 97%

Section: Introductionmentioning

confidence: 99%

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Metabolism of 3′-Azido-3′-Deoxythymidine and 3′-Fluoro-3′-Deoxythymidine in Combination in Human Immunodeficiency Virus Infected Lymphoblastoid Cells

Cox

1992

Antivir Chem Chemother

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“…In particular, modifications of3' substituent on the sugar moiety are used, due to the fact that this may cause a chain termination in the synthesis ofviral nucleic acids. Thymidine analogues have proved to be effective as antiretroviral agents, and the in vitro and in vivo antiviral effects of alovudine (3'-fluorothymidine) and zidovudine (:'r-azidothymidine) have been well documented by Mitsuya et al (1985); Matthes et al (1987Matthes et al ( , 1988 ;Good and de Miranda (1992); and Sandstrom and Oberg (1993). Their pharmacokinetic properties have also been studied in detail (Balis et al, 1989;Boudinot et al, 1990;Good and de Miranda, 1992;Ljungdahl-Stahle et al, 1992;Sandstrom and Oberg, 1993;Stahle et al, 1993).…”

Section: Introductionmentioning

confidence: 99%

Distribution to the Brain and Protein Binding of 3′and 5-Substituted 2′,3′-Dideoxyuridine Derivatives, Studied by Microdialysis

Borg

Zhou

Johansson

et al. 1997

Antivir Chem Chemother

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SummaryThe aim of this study was to investigate a series of 3' and 5-substituted 2',3' -dideoxyuridine derivatives (ddUD) with respect to plasma protein binding, halflife and distribution across the blood-brain barrier in the rat. The microdialysis technique was used to study protein binding in human plasma (in vitro), and to sample the extracellular space of rats with microdialysis probes implanted into the striatum of the brain and the gastrocnemic muscle (in vivo). The compounds were analysed by HPLC with UV-detection. The octanol/water partition coefficients of the dduD varied from 0.08-0.84. The protein binding of the ddUDswas approximately 80%. After s.c. administration (25 or 50 mg kg-1 ), the brain and muscle extracellular levels differed; brain levels were 0.18-0.36 o] peripheral (muscle) concentrations. A multivariate analysis, which included data on zidovudine, alovudine and thymidine, demonstrated a relationship between the physicochemical and some of the phormacokinetic properties of uridine analogues. The analysis shows that half-life and protein binding increases with decreasing pK a . However, penetration to the brain is not correlated with the partition into octano!. It is concluded that the transport to the brain is not primarily dependent upon passive diffusion over a lipophilic barrier but, rather, to other chemical properties of the dduDs. This is suggestive of a specific transport mechanism, e.g. the thymidine carrier.

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“…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) are potent inhibitors of endogenous HBV DNA polymerase (14; E. Matthes In vitro assay for antiviral activity.…”

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confidence: 99%

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

Matthes

Langen

Janta-Lipinski

et al. 1990

Antimicrob Agents Chemother

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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...

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Phosphorylation, anti-HIV activity and cytotoxicity of 3′-fluorothymidine

Cited by 83 publications

References 10 publications

Metabolism of 3′-Azido-3′-Deoxythymidine and 3′-Fluoro-3′-Deoxythymidine in Combination in Human Immunodeficiency Virus Infected Lymphoblastoid Cells

Metabolism of 3′-Azido-3′-Deoxythymidine and 3′-Fluoro-3′-Deoxythymidine in Combination in Human Immunodeficiency Virus Infected Lymphoblastoid Cells

Distribution to the Brain and Protein Binding of 3′and 5-Substituted 2′,3′-Dideoxyuridine Derivatives, Studied by Microdialysis

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

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