Inhibition of HIV-associated reverse transcriptase by sugar-modified derivatives of thymidine 5′-triphosphate in comparison to cellular DNA polymerases α and β

Matthes, E.; Lehmann, Christian; Scholz, D.; Janta-Lipinski, M. Von; Gaertner, K.; Rosenthal, H. A.; Langen, P.

doi:10.1016/0006-291x(87)91078-3

Cited by 101 publications

(63 citation statements)

References 8 publications

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“…We and others have found previously that AZT-TP inhibits the RNA-directed DNA polymerase activity of HIV-1 reverse transcriptase by reversibly competing with dTTP (Furmanet al, 1986;Matthes et al, 1987;St. Clair et al, 1987;Parker et al, 1991).…”

Section: Kinetic Analysis Of the Anti-reverse Transcriptase Synergy Bmentioning

confidence: 93%

Cell-Based and Biochemical Analysis of the anti-HIV Activity of Combinations of 3′-azido-3′-deoxythymidine and Analogues of TIBO

Buckheit

White

Germany-Decker

et al. 1994

Antivir Chem Chemother

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The toxicity of 3′-azido-3′-deoxythymidine (AZT) and the appearance of drug-resistant mutants in patients treated with AZT emphasizes the critical importance of the development of alternative strategies for the therapy of AIDS patients. Combination antiviral chemotherapy provides an attractive therapeutic strategy since the dose of the individual agents may be lowered to reduce toxicity and the use of two potent antiviral agents may limit the development of drug resistance. Two analogues of tetrahydro-imidazo[4,5,1-jk][1,4]-benzodiazepin-2(1H)-thione (TIBO) potently and selectively inhibit the replication of HIV-1 in cell culture. In combination with AZT, either of the two TIBO compounds, R82913 and R86183, was highly synergistic in cell culture against HIV-1. However, in biochemical enzyme inhibition assays, utilizing recombinant HIV-1 reverse transcriptase, synergy was not detected at the enzymatic level. These results suggest that one of these two known inhibitors of HIV-1 reverse transcriptase may have a secondary mechanism of action distinct from inhibition of the reverse transcriptase.

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Section: Kinetic Analysis Of the Anti-reverse Transcriptase Synergy Bmentioning

confidence: 93%

Cell-Based and Biochemical Analysis of the anti-HIV Activity of Combinations of 3′-azido-3′-deoxythymidine and Analogues of TIBO

Buckheit

White

Germany-Decker

et al. 1994

Antivir Chem Chemother

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“…In the literature, the mechanism by which AZT and dideoxynucleosides (as their 5'-triphosphates) inhibit HIV is reported to be inhibition of retroviral reverse transcriptase (7,15,25,28,36,41). Reverse transcriptase inhibition has become a commonly used biological endpoint for the discovery of new anti-HIV agents (7,25,36,41).…”

Section: Resultsmentioning

confidence: 99%

“…A proposed mechanism of action of these agents is the inhibition of human immunodeficiency virus (HIV) reverse transcriptase after nucleoside phosphorylation to their respective 5'-triphosphates (7,15,18,25,28,36,41). These agents have other biological activities, notably, an ability to terminate DNA chain elongation (1, 38, 39).…”

mentioning

confidence: 99%

Induction of the SOS response in Escherichia coli by azidothymidine and dideoxynucleosides

Mamber

Brookshire

Forenza

1990

Antimicrob Agents Chemother

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A number of nucleosides with anti-human immunodeficiency virus (HIV) activity were evaluated in two colorimetric (j-galactosidase) assays for induction of the SOS response in Escherichia coli. 3'-Azido-3'-deoxythymidine (azidothymidine; AZT), 2',3'-dideoxyadenosine (ddA), 2',3'-dideoxyguanosine (ddG), and 2',3'-dideoxyinosine (ddl) induced cel filamentation (sulA) and prophage lambda in well-agar diffusion and liquid microsuspension assays. AZT was approximately 100 times more potent than the dideoxypurine nucleosides, inducing sulA at <100 ng/ml. 2',3'-Dideoxythymidine (ddT) and 2',3'-dideoxy-2',3'-didehydrothymidine (D4T) induced sulA at 100 to 1,000 jg/ml, while 2',3'-dideoxycytidine (ddC) weakly induced prophage lambda. Activity relationships thus were AZT > ddA 2 ddl 2 ddG > ddT = D4T > ddC. ddA and ddl had equivalent activities in agar diffusion assays, but different activity profiles were observed in liquid microsuspension assays. The differences may be related to drug metabolism. AZT and ddA showed marginal effects in a DNA repair (preferential toxicity) assay in which E. coli WP2 and CM871 uvrA recA lexA were used. Furthermore, none of the agents was able to preferentially inhibit BaciUus subtilis M45 recA relative to wild-type strain H17. These data suggest that AZT and the dideoxynucleosides do not cause DNA lesions that are repairable by excision repair and/or error-free postreplication repair processes. Rather, the SOS response appears to be induced by DNA chain termination leading to the inhibition of DNA replication. Bacterial assays for induction of the SOS response may be useful as simple, rapid prescreens for the discovery of new anti-HIV agents. Moreover, such assays may provide an additional parameter in the evaluation of agents with demonstrated activity against HIV and other retroviruses.3'-Azido-3'-deoxythymidine (azidothymidine; AZT) and dideoxynucleosides such as 2',3'-dideoxyadenosine (ddA) and 2',3'-dideoxyinosine (ddI) have potential value in the chemotherapy of patients with acquired immunodeficiency syndrome (3,4,26,28,47). A proposed mechanism of action of these agents is the inhibition of human immunodeficiency virus (HIV) reverse transcriptase after nucleoside phosphorylation to their respective 5'-triphosphates (7,15,18,25,28,36,41). These agents have other biological activities, notably, an ability to terminate DNA chain elongation (1, 38, 39). DNA chain termination may play a role in the effects of these agents on HIV (2,4,14,26,27,36). Moreover, DNA chain termination is the mechanism postulated to explain the bactericidal effects of ddA and AZT against Escherichia coli and other members of the family Enterobacteriaceae (6,13). In 1972, Geissler et al. (16) reported that ddA induces prophage lambda in E. coli. Elwell et al. (13) noted that AZT causes filamentation of E. coli cells. These results suggest that AZT and the dideoxynucleosides are capable of inducing the SOS response (21, 34, 42-44), either by causing DNA damage or by inhibiting DNA replication. However, in the Sa...

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“…Because the cytostatic or cytotoxic effects of the compounds were measured by two independent methods, incorporation inhibition studies and direct cell counting, it appears to be unlikely that the antiviral effect that we observed was due to an inhibition of cellular protein synthesis. The high antiviral selectivity could be explained by the high affinity of the triphosphate of FddThd as a competitive inhibitor toward different DNA polymerases (12). 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.…”

Section: Discussionmentioning

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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Inhibition of HIV-associated reverse transcriptase by sugar-modified derivatives of thymidine 5′-triphosphate in comparison to cellular DNA polymerases α and β

Cited by 101 publications

References 8 publications

Cell-Based and Biochemical Analysis of the anti-HIV Activity of Combinations of 3′-azido-3′-deoxythymidine and Analogues of TIBO

Cell-Based and Biochemical Analysis of the anti-HIV Activity of Combinations of 3′-azido-3′-deoxythymidine and Analogues of TIBO

Induction of the SOS response in Escherichia coli by azidothymidine and dideoxynucleosides

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

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