5-ethyl-2′-deoxyuridine: Evidence for incorporation into DNA and evaluation of biological properties in lymphocyte cultures grown under conditions of amethopterine-imposed thymidine deficiency

Swierkowska, Krystyna; Jasińska, J; Steffen, Jan

doi:10.1016/0006-2952(73)90257-8

Cited by 21 publications

(7 citation statements)

References 8 publications

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“…This communication reports the results of a comparative study on the antiviral and cytotoxic effects of all four analogues and suggests that selective affinity to various TK's is responsible for their differential activity. Some studies on the biological activity of 5ethyl-, 5-vinyl-, and 5-allyl-dUrd have been reported previously by other investigators (2,7,(9)(10)(11).…”

mentioning

confidence: 74%

Antiviral Action and Cellular Toxicity of Four Thymidine Analogues: 5-Ethyl-, 5-Vinyl-, 5-Propyl-, and 5-Allyl-2′-Deoxyuridine

Cheng

Domin

Sharma

et al. 1976

Antimicrob Agents Chemother

106

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5-Ethyl-, 5-vinyl-, 5-propyl-, and 5-allyl-2'-deoxyuridine (dUrd) had antiviral activity against herpes simplex type 1 and type 2 grown in HeLa TK-cells, in the order 5-vinyl-dUrd, 5-ethyl-dUrd, 5-propyl-dUrd, 5-allyl-dUrd, but they were inactive against a TK-mutant of herpes simplex type 1. The antiviral activity of these compounds could be partially reversed by thymidine. Except for 5-vinyl-dUrd, they were not toxic to WI-38 and HeLa TK-cells at a concentration of 25 ,uM. All four analogues inhibited the growth of herpes simplex type 1-transformed HeLa TK-cells at a concentration of 1 ,uM.Herpesviruses are responsible for many diseases in man, and recent evidence also suggests a strong correlation between herpes infections and the incidence of certain human malignancies (4). One characteristic of herpes simplex type I (HSV-1) and type II (HSV-2) is that they can induce virus-specific thymidine kinases (TK) in some human cells after infection. These viral TK's are different from the cytoplasmic and mitochondrial TK's of human cells not only with respect to their physical and chemical properties, such as electrophoretic mobility, activation energy, and salt sensitivity, but also in substrate specificity (Y. C. Cheng, submitted for publication; L. S. Lee and Y. C. Cheng, submitted for publication). This fact suggested to us that specific anti-HSV-1 and HSV-2 agents that specifically utilize viral TK might be developed.The approach taken in this laboratory is to seek analogues of thymidine that could act as alternative substrates for viral TK Chem., in press). A previous study on the binding specificity of some pyrimidine 2'-deoxyribonucleosides showed that 5-ethyl-, 5-propyl-, and 5-allyl-deoxyuridine (dUrd) had good binding affinity to HSV-1-and HSV-2-specific TK's (Km of thymidine to Ki of analogue less than 0.2) but much poorer affinity for either human cytosol or mitochondrial TK's (Km of thymidine to K, of analogue larger than 0.005) and that 5-vinyl-dUrd had a high binding affinity to both viral and human mitochondrial TK's. For instance, the Ki of 5-ethyl-dUrd for human cytosol, human mitochondrial, HSV-1 (strain KOS), and HSV-2 (strain 333) TK's was 82, 305, 0.7, and 0.3 ,uM, respectively. TheKi of 5-vinyl-dUrd for human cytosol, human mitochondrial, HSV-1 (strain KOS), and HSV-2 (strain 333) TK's was 35, 1.7, 0.5, and 0.5 ,iM, respectively (Cheng and Lee, submitted for publication). This communication reports the results of a comparative study on the antiviral and cytotoxic effects of all four analogues and suggests that selective affinity to various TK's is responsible for their differential activity. Some studies on the biological activity of 5-ethyl-, 5-vinyl-, and 5-allyl-dUrd have been reported previously by other investigators (2, 7, 9-11).MATERIALS AND METHODS Cells and viruses. The stock viruses, HSV-1 (strains KOS and B2006) and HSV-2 (strain 333), were kindly given to us by W. Munyon of this institute (1, 3). Both the KOS and 333 strains of herpes simplex viruses but not the B2006 strain cou...

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confidence: 74%

Antiviral Action and Cellular Toxicity of Four Thymidine Analogues: 5-Ethyl-, 5-Vinyl-, 5-Propyl-, and 5-Allyl-2′-Deoxyuridine

Cheng

Domin

Sharma

et al. 1976

Antimicrob Agents Chemother

106

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“…Historically, the first method involved the incorporation of [ 3 H]-thymidine into DNA during S-phase of the cell cycle (Swierkowska et al, 1973). This method is very useful for quantitative analyses of cell division but has shortcomings due to the labour-intensive and time-consuming processing required for detecting the radioactivity (Taupin, 2007).…”

Section: Discussionmentioning

confidence: 99%

EdU, a new thymidine analogue for labelling proliferating cells in the nervous system

Chehrehasa

Meedeniya

Dwyer

et al. 2009

Journal of Neuroscience Methods

248

189

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Labelling and identifying proliferating cells is central to understanding neurogenesis and neural lineages in vivo and in vitro. We present here a novel thymidine analogue, ethynyl deoxyuridine (EdU) for labelling dividing cells, detected with a fluorescent azide which forms a covalent bond via the "click" chemistry reaction (the Huisgen 1,3-dipolar cycloaddition reaction of an organic azide to a terminal acetylene). Unlike the commonly used BrdU, EdU detection requires no heat or acid treatment. It is quick and easy and compatible with multiple probes for fluorescence immunochemistry, facilitating the characterisation of proliferating cells at high resolution.

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Section: -Alkyl Nucleosidesmentioning

confidence: 99%

“…In reversions of T4 rI1 mutants to r+, where BUdR gives the expected frequencies of revertants, EtUdR was without effect [27]. EtUdR restores cell proliferation in human phytohemagglutinin-stimulated lymphocytes grown under condi-tions of DNA synthesis block due to amethopterine induced thymidine deficiency, with incorporation into nuclei of cells entering the S phase; the analogue induced no chromatid aberrations in metaphase under conditions where BUdR did so [45]. EtUdR was found to be almost as effective as IUdR and BUdR against vaccinia virus on HeLa cells [46] , and was somewhat less effective than IUdR against herpes simplex on chick embryo fibroblast monolayers (H. Kaufman, personal communication).…”

Section: -Alkyl Nucleosidesmentioning

confidence: 99%

Progress with antiviral agents

Shugar

1974

FEBS Letters

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5-ethyl-2′-deoxyuridine: Evidence for incorporation into DNA and evaluation of biological properties in lymphocyte cultures grown under conditions of amethopterine-imposed thymidine deficiency

Cited by 21 publications

References 8 publications

Antiviral Action and Cellular Toxicity of Four Thymidine Analogues: 5-Ethyl-, 5-Vinyl-, 5-Propyl-, and 5-Allyl-2′-Deoxyuridine

Antiviral Action and Cellular Toxicity of Four Thymidine Analogues: 5-Ethyl-, 5-Vinyl-, 5-Propyl-, and 5-Allyl-2′-Deoxyuridine

EdU, a new thymidine analogue for labelling proliferating cells in the nervous system

Progress with antiviral agents

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