A comparison of the enantioselectivities of human deoxycytidine kinase and human cytidine deaminase∗

Shafiee, Manijeh; Griffon, Jean-François; Gosselin, Gilles; Cambi, Alessandra; Vincenzetti, Silvia; Vita, Alberto; Eriksson, Staffan; Imbach, Jean‐Louis; Maury, Georges

doi:10.1016/s0006-2952(98)00165-8

Cited by 17 publications

(12 citation statements)

References 40 publications

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“…This enzymatic reaction was described as one main metabolic pathway of the D-stereoisomer D-HyCR (8). However, we found that L-Hyd4C is completely resistant against deamination, confirming the described strict enantioselectivity of this enzyme (18). The other main pathway described for D-HyCR is a reduction of the NHOH group to the NH 2 group, giving D-cytidine.…”

Section: Discussionsupporting

confidence: 81%

“…Human deoxycytidine kinase has been described earlier as having only a low enantioselectivity, which also favors the phosphorylation of a series of L-cytidine derivatives, such as 3TC (2), its 5-fluoro derivative (FTC) (14), L-ddC and its 5-fluoro derivative (L-ddFC), and L-CdR (11,18). Ten M L-Hyd4C was converted to 11.5 Ϯ 1.8 pmol monophosphate/ 10 6 cells during 24 h.…”

Section: Discussionmentioning

confidence: 99%

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Cellular Pharmacology of the Anti-Hepatitis B Virus Agent β- l -2′,3′-Didehydro-2′,3′-Dideoxy-N ⁴ -Hydroxycytidine: Relevance for Activation in HepG2 Cells

Matthes

Bünger

2010

Antimicrob Agents Chemother

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A large number of nucleoside analogues have been described as inhibitors of hepatitis B virus (HBV) and HIV replication. Recently L-nucleoside analogues in particular have gained increasing interest. They are characterized by an opposite configuration from that of the natural D-nucleoside analogues and represent one of the most attractive groups of antiretroviral compounds, including ß-L-2Ј,3Ј-dideoxy-3-thiacytidine (3TC) and its, and ß-L-2Ј,3Ј-didehydro-2Ј,3Ј-dideoxy-2Ј-fluoro-cytidine (L-2ЈFd4C) (3,5,22).Some of them not only have been found to be more potent than their corresponding D-nucleosides but seem to exhibit lower cytotoxicity and have been proved to be effective and selective agents for the treatment of chronic hepatitis B virus infections (4). However, only long-term therapy with a single nucleoside for several years was shown to be able to completely suppress HBV DNA in serum of patients and to reverse the progression of the disease. The disadvantage connected with such therapy regimens is the development of drug-resistant HBV strains (22). Therefore, the challenge will be to develop more-efficient drugs for shorter treatment regimens and to combine them to reach synergistic or at least additive drug action. This approach has been described not only as being highly efficient for the treatment of HIV infections but also as preventing the development of resistant mutants. Therefore, AIDS therapy is considered a model for future therapy of chronic HBV infections (17).Recently we described a series of new ß-L-N 4 -hydroxydeoxycytidine and ß-L-5-methyl-deoxycytidine derivatives as inhibitors of HBV replication. Between them, ß-L-2Ј,3Ј-didehydro-2Ј,3Ј-dideoxy-N 4 -hydroxycytidine (L-Hyd4C) (Fig. 1) emerged as the most effective in suppression of virus production in HepG2.2.15 cells (50% effective dose [ED 50 ] ϭ 0.03 M), displaying an extremely low cytotoxicity (50% cytotoxic dose [CD 50 ] for HepG2 cells ϭ 2,500 M) (12).These encouraging features have prompted us to investigate the cellular pharmacology of L-Hyd4C in a hepatic cell line. This included the activation of this unnatural L-deoxycytidine nucleoside to its 5Ј-mono-, 5Ј-di-, and 5Ј-triphosphate, the search for other metabolites, and the estimation of the intracellular halflives (t 1/2 ) of the 5Ј-di-and 5Ј-triphosphate of (This work was presented in part at BIT's 5th Anniversary Congress of International Drug Discovery Science and Technology, 7 to 13 November 2007, Xi'an and Beijing, China.) MATERIALS AND METHODSCompounds. The synthesis and characterization of L-Hyd4C and its 5Ј-triphosphate were described elsewhere (E. Matthes, M. von Janta-Lipinski, H. Will, H. Sirma, and A. High-performance liquid chromatography (HPLC) analysis by methods described below showed 12% of radioactive contamination of the tritiated products which were separated by HPLC.Acetonitrile (gradient grade), tetrabutylammonium phosphate (monobasic, purissimum), and 3-deazauridine (deazaUR) were obtained from Sigma-Aldrich (Taufkirchen, Germany). Ammonium dihydrogen ph...

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Section: Discussionsupporting

confidence: 81%

Section: Discussionmentioning

confidence: 99%

Cellular Pharmacology of the Anti-Hepatitis B Virus Agent β- l -2′,3′-Didehydro-2′,3′-Dideoxy-N ⁴ -Hydroxycytidine: Relevance for Activation in HepG2 Cells

Matthes

Bünger

2010

Antimicrob Agents Chemother

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“…Viral polymerases incorporate l -derivatives more readily than do the human ones (20–21). Nucleoside degrading enzymes, such as nucleoside deaminases and phosphorylases, tend to be strictly selective for d -nucleotides although only limited data is available (11,22). The lack of degradation of l -nucleosides has however been demonstrated in mice, where l -nucleosides were gradually excreted in the unchanged form following intraperitoneal administration (5).…”

Section: Introductionmentioning

confidence: 99%

Enantioselectivity of human AMP, dTMP and UMP-CMP kinases

Alexandre¹,

Roy²,

Topalis³

et al. 2007

Nucleic Acids Research

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l-Nucleoside analogues such as lamivudine are active for treating viral infections. Like d-nucleosides, the biological activity of the l-enantiomers requires their stepwise phosphorylation by cellular or viral kinases to give the triphosphate. The enantioselectivity of NMP kinases has not been thoroughly studied, unlike that of deoxyribonucleoside kinases. We have therefore investigated the capacity of l-enantiomers of some natural (d)NMP to act as substrates for the recombinant forms of human uridylate-cytidylate kinase, thymidylate kinase and adenylate kinases 1 and 2. Both cytosolic and mitochondrial adenylate kinases were strictly enantioselective, as they phosphorylated only d-(d)AMP. l-dTMP was a substrate for thymidylate kinase, but with an efficiency 150-fold less than d-dTMP. Both l-dUMP and l-(d)CMP were phosphorylated by UMP-CMP kinase although much less efficiently than their natural counterparts. The stereopreference was conserved with the 2′-azido derivatives of dUMP and dUMP while, unexpectedly, the 2′-azido-d-dCMP was a 4-fold better substrate for UMP-CMP kinase than was CMP. Docking simulations showed that the small differences in the binding of d-(d)NMP to their respective kinases could account for the differences in interactions of the l-isomers with the enzymes. This in vitro information was then used to develop the in vivo activation pathway for l-dT.

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“…As expected from the similarities in the structures of the active sites of the two deaminases, the enantioselectivity of CDA follows the same trend as that of ADA. Thus, the β-L-enantiomer of dC, or the α-L-and β-L-enantiomers of riboC, araC, xyloC and lyxoC were resistant to human CDA (Shafiee et al, 1998). Similarly, the β-L-enantiomers of 3TC (Chang et al, 1992a), FTC (Furman et al, 1992), FddC (Martin et al, 1997) and Fd4C were not substrates or weak substrates of CDA, contrary to the corresponding β-D-enantiomers.…”

Section: Enantioselectivities Of Nucleoside Deaminases and Phosphorylmentioning

confidence: 99%

The Enantioselectivity of Enzymes Involved in Current Antiviral Therapy Using Nucleoside Analogues: A New Strategy?

Maury

2000

Antivir Chem Chemother

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This review is primarily intended for synthetic bio-organic chemists and enzymologists who are interested in new strategies in the design of virus inhibitors. It is an attempt to assess the importance of the enzymatic properties of L-nucleosides and their analogues, particularly those that are active against viruses such as human immunodeficiency virus (HIV), hepatitis B virus (HBV), herpes simplex virus (HSV), etc. Only data obtained with purified enzymes have been considered and discussed. The examined enzymes include nucleoside- or nucleotide-phosphorylating enzymes, catabolic enzymes, viral target enzymes and cellular polymerases. The enantioselectivities of these enzymes were determined from existing data and are significant only when a sufficient number of enantiomeric pairs of substrates could be examined. The reported data emphasize the weak enantioselectivities of cellular or viral nucleoside kinases and some viral DNA polymerases. Thus, cellular deoxycytidine kinase has a considerably relaxed enantioselectivity with respect to a large number of nucleosides or their analogues, and it occupies a strategic position in the intracellular activation of the compounds. Similarly, HIV-1 reverse transcriptase often has a relatively weak enantioselectivity and can be inhibited by the 5-triphosphates of a large series of L-nucleosides and analogues. In contrast, degradation enzymes, such as adenosine or cytidine deaminases, generally demonstrate strict enantioselectivities favouring D-enantiomers and are used by chemists in asymmetric syntheses. The weak enantioselectivities of some enzymes involved in nucleoside metabolism are more or less pronounced, and one enantiomer or the other is favoured depending on the substrate. This suggests that the low enantioselectivity is fortuitous and does not result from evolutionary pressure, since these enzymes do not create or modify asymmetric centres in substrates. The combined enantioselectivities of the enzymes examined in this review strongly suggest that the field of L-nucleosides and their analogues should be systematically explored in the search for new virus inhibitors.

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A comparison of the enantioselectivities of human deoxycytidine kinase and human cytidine deaminase∗

Cited by 17 publications

References 40 publications

Cellular Pharmacology of the Anti-Hepatitis B Virus Agent β- l -2′,3′-Didehydro-2′,3′-Dideoxy-N ⁴ -Hydroxycytidine: Relevance for Activation in HepG2 Cells

Cellular Pharmacology of the Anti-Hepatitis B Virus Agent β- l -2′,3′-Didehydro-2′,3′-Dideoxy-N ⁴ -Hydroxycytidine: Relevance for Activation in HepG2 Cells

Enantioselectivity of human AMP, dTMP and UMP-CMP kinases

The Enantioselectivity of Enzymes Involved in Current Antiviral Therapy Using Nucleoside Analogues: A New Strategy?

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A comparison of the enantioselectivities of human deoxycytidine kinase and human cytidine deaminase∗

Cited by 17 publications

References 40 publications

Cellular Pharmacology of the Anti-Hepatitis B Virus Agent β- l -2′,3′-Didehydro-2′,3′-Dideoxy-N 4 -Hydroxycytidine: Relevance for Activation in HepG2 Cells

Cellular Pharmacology of the Anti-Hepatitis B Virus Agent β- l -2′,3′-Didehydro-2′,3′-Dideoxy-N 4 -Hydroxycytidine: Relevance for Activation in HepG2 Cells

Enantioselectivity of human AMP, dTMP and UMP-CMP kinases

The Enantioselectivity of Enzymes Involved in Current Antiviral Therapy Using Nucleoside Analogues: A New Strategy?

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Cellular Pharmacology of the Anti-Hepatitis B Virus Agent β- l -2′,3′-Didehydro-2′,3′-Dideoxy-N ⁴ -Hydroxycytidine: Relevance for Activation in HepG2 Cells

Cellular Pharmacology of the Anti-Hepatitis B Virus Agent β- l -2′,3′-Didehydro-2′,3′-Dideoxy-N ⁴ -Hydroxycytidine: Relevance for Activation in HepG2 Cells