L-Thymidine is phosphorylated by herpes simplex virus type 1 thymidine kinase and inhibits viral growth

Spadari, Silvio; Maga, Giovanni; Focher, Federico; Ciarrocchi, Giovanni; Manservigi, Roberto; Arcamone, Federico; Capobianco, Massimo L.; Carcuro, Antonio; Colonna, Francesco Paolo

doi:10.1021/jm00100a029

Cited by 106 publications

(67 citation statements)

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“…To this purpose, chemical modifications of the sugar ring, such as replacement of the 4h-oxygen by a methylene group [1,2] or replacement of the -2h-deoxyribose or the carbocyclic -2h-deoxyribose by their respective  enantiomers [3][4][5] have indeed given rise to nucleosides resistant to such degradation and which remain active as anticancer and antiviral agents [17]. Replacement of the 4h-oxygen by sulphur was also studied, but only in the case of ribonucleosides [18].…”

Section: Discussionmentioning

confidence: 99%

“…HeLa TK − cells (human epithelial cell line derived from a cervix carcinoma, TK-deficient) and HeLa TK − \HSV-1 TK + (HeLa TK − transformed to the TK + phenotype with a functional copy of the HSV-1 TK gene) were those previously used [3]. Cells were tested for mycoplasma contamination with the Hoechst 33258 staining method and were found to be negative.…”

Section: Cells and Virus Assaysmentioning

confidence: 99%

“…In an attempt to avoid this degradation, modified nucleosides, such as carbocyclic derivatives [1,2], have been synthesized in which the sugar moiety has been replaced by a cyclopentyl or a cyclobutyl ring, or enantiomeric derivatives [3,4], in which the -deoxyribose moiety has been replaced by the  enantiomer [3][4][5][6]. More recently, several 4h-thio-2h-deoxyribonucleosides have been synthesized and studied for their possible antiviral activity [7].…”

Section: Introductionmentioning

confidence: 99%

“…2 Present address : Melford Laboratories Ltd., Bildeston Rd. Chelsworth, Ipswich, Suffolk IP7 7LE, U.K. 3 Deceased. 4 To whom correspondence should be sent (e-mail spadari!igbe.pv.cnr.it).…”

Section: Introductionmentioning

confidence: 99%

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Anti-(herpes simplex virus) activity of 4′-thio-2′-deoxyuridines: a biochemical investigation for viral and cellular target enzymes

Verri

Focher

Duncombe

et al. 2000

Biochemical Journal

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The antiviral activity of several nucleoside analogues is often limited by their rapid degradation by pyrimidine nucleoside phosphorylases. In an attempt to avoid this degradation, several modified nucleosides have been synthesized. A series of 4h-thio2h-deoxyuridines exhibits an anti-[herpes simplex virus (HSV)] activity significantly higher (20-600 times) than that shown by the corresponding 4h-oxy counterpart. We investigated the mode of action of these compounds and we found that : (i) several 4h-thio-2h-deoxyuridines are phosphorylated to the mono-and diphosphates by HSV-1 thymidine kinase (TK) more efficiently than their corresponding 4h-oxy counterpart ; (ii) both are inhibitors of cellular thymidylate synthase ; (iii) 4h-thio-2h-deoxyuridines are resistant to phosphorolysis by human thymidine phosphorylase ; (iv) both 4h-oxy-and 4h-thio-2h-deoxyuridines are phosphorylated to deoxyribonucleotide triphosphate in HSV-1-infected cells and are incorporated into viral DNA ; (v) 4h-thio-

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Section: Discussionmentioning

confidence: 99%

Section: Cells and Virus Assaysmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Anti-(herpes simplex virus) activity of 4′-thio-2′-deoxyuridines: a biochemical investigation for viral and cellular target enzymes

Verri

Focher

Duncombe

et al. 2000

Biochemical Journal

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“…Based on the lack of enantioselectivity of HSV-TK in comparison with mammalian TK, 14 we used L-thymidine (LT), the enantiomer of the naturally occurring D-thymidine, in order to trace HSV-TK-containing cells without interfering with cell metabolism and survival ( Figure 1). LT is considered very safe, and neither in vitro nor in vivo studies have shown any sign of cellular or mitochondrial toxicity.…”

Section: Introductionmentioning

confidence: 99%

Lack of enantioselectivity of herpes virus thymidine kinase allows safer imaging of gene delivery

et al. 2003

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Herpes simplex virus thymidine kinase (HSV-TK) is widely used in gene therapy. The enzymatic activity of HSV-TK may be traced in vivo by specific radiopharmaceuticals in order to image transgene expression. However, most of these radiopharmaceuticals are toxic per se or after activation by HSV-TK, and therefore do not represent ideal molecules for clinical applications and repeated imaging. Unlike human cytosolic TK, HSV-TK is not enantioselective and can efficiently phosphorylate both D and L enantiomers of b-thymidine. Here we show that, after phosphorylation by HSV-TK, tritiated L-b-thymidine (LT) is selectively retained inside the cells in vitro and in vivo. We used the in vivo accumulation of radioactive phosphorylated LT to image the HSV-TK-positive cells inside a transplantable murine brain tumour after inoculation of cells producing retroviruses carrying HSV-TK. Owing to their unnatural enantiomeric conformation, phosphorylated LT metabolites are very poorly processed by mammalian enzymes, thus leading to increased cellular retention and minimal toxicity. The ability to image cells expressing the HSV-TK gene by using radiolabelled LT, without damaging the cells accumulating the phosphorylated L-nucleoside, will be important to monitor the levels and spatial distribution of therapeutic vectors carrying HSV-TK.

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Synthesis Of Nucleosides

Vorbrüggen¹,

1999

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This chapter deals with the synthesis of nucleosides (e.g., the formation of N ‐glycosides of sugars such as D ‐ribose or 2‐deoxy‐D‐ribose with heterocyclic nitrogen bases). The methods of nucleoside synthesis have been treated in a number of reviews and monographs. It is now generally accepted that nucleosides were among the first organic compounds formed at the start of evolution in the early history of our planet earth. To support this point, guanine and adenine were heated with D ‐ribose in seawater, which contains the Lewis acid magnesium chloride as catalyst. One thus obtained the nucleosides guanosine and adenosine together with comparable yields of unnatural α‐nucleosides. The latter were gradually photoanomerized to the thermodynamically more stable compounds in overall yields of 5–6%. The furanose form of ribose reacts faster than the pyranose form. Corresponding syntheses of the pyrimidine nucleosides uridine and cytidine from uracil , cytosine and ribose are more problematic and remain an enigma. The recent conversion of glycolaldehyde‐ O ‐phosphate and formaldehyde to ribose‐2,4‐di‐ O ‐phosphate might give new insights into the prebiotic syntheses of uridine and cytidine. The evidence and hypotheses for these prebiotic conversions and the evolution of RNA, as well as the implications of an “RNA World,” have been reviewed. These RNA nucleosides are reduced in vivo as 5′‐ O ‐diphosphates by ribonucleotide reductases to the corresponding 2′‐deoxynucleosides—the building blocks of DNA such as 2′‐deoxyguanosine. The thermodynamically controlled synthesis of these four building blocks of RNA has implications for the design of efficient, high yielding, new methods for the synthesis of the naturally occurring nucleosides, nucleoside antibiotics, and modified nucleosides that may serve as antimetabolites to fight viral and parasitic diseases and cancer. The nucleoside rings in this chapter are depicted arbitrarily in the anti conformation, as occurs predominantly in the crystal and solution (based primarily on NOE‐ 1 H‐ and 13 C‐NMR measurements) forms of pyrimidine nucleosides. Only a few nucleosides, such as 6‐methyluridine, occur with the heterocyclic ring predominantly in the syn conformation. The synthesis of C ‐nucleosides has been reviewed previously and is not covered in this review.

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L-Thymidine is phosphorylated by herpes simplex virus type 1 thymidine kinase and inhibits viral growth

Cited by 106 publications

References 1 publication

Anti-(herpes simplex virus) activity of 4′-thio-2′-deoxyuridines: a biochemical investigation for viral and cellular target enzymes

Anti-(herpes simplex virus) activity of 4′-thio-2′-deoxyuridines: a biochemical investigation for viral and cellular target enzymes

Lack of enantioselectivity of herpes virus thymidine kinase allows safer imaging of gene delivery

Synthesis Of Nucleosides

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