RESUMO
A series of 5-substituted 2'-deoxy-4'-thiopyrimidine nucleosides was synthesized and evaluated as potential antiviral agents. A number of analogues such as 2'-deoxy-5-propyl-4'-thiouridine (3ii), 2'-deoxy-5-isopropyl-4'-thiouridine (3iii), 5-cyclopropyl-2'-deoxy-4'-thiouridine (3iv), 2'-deoxy-4'-thio-5-vinyluridine (3viii), and 5-(2-chloroethyl)-2'-deoxy-4'-thiouridine (3xx) were found to be highly active against herpes simplex virus type-1 (HSV-1) and varicella zoster virus (VZV) in vitro with no significant cytotoxicity. The compound with the broadest spectrum of activity was 2'-deoxy-5-ethyl-4'-thiouridine (3i) which showed significant activity against HSV-1, HSV-2, and VZV.
Assuntos
Antivirais/síntese química , Antivirais/farmacologia , Nucleosídeos de Pirimidina/síntese química , Nucleosídeos de Pirimidina/farmacologia , Simplexvirus/efeitos dos fármacos , Animais , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Chlorocebus aethiops , Humanos , Células Vero , Ensaio de Placa ViralRESUMO
Derivatives of such related substances as cytosine, uracil, thymine, 6-methyl-uracil, 5-ethyluracil, 5-propyluracil, 5-isopropyluracil, 5-cyclopropyluracil, 5-allyluracil, 5,6-trimethyleneuracil, 6-cyclopropyluracil, 5-cyclobutyluracil and 5-tert-butyluracil have been separated on a column of Spheron P-300. Retention on the column was found to depend on the size of the non-polar part of the molecule. The chromatographic behaviour is analyzed according to the theory of solvophobic chromatography.
Assuntos
Uracila/análogos & derivados , Fenômenos Químicos , Química , Cromatografia Líquida/instrumentação , Cromatografia Líquida/métodos , Pirimidinas/isolamento & purificação , Solubilidade , Temperatura , Uracila/isolamento & purificaçãoRESUMO
A series of 5-substituted pyrimidine 4'-thio-2'-deoxynucleosides has been synthesized and their antiviral properties determined. It seems likely that once an analogue is a kinase substrate, then that analogue is toxic for that organism. Thus 4'-thiothymidine is phosphorylated by both viral and host kinases and shows toxicity to viruses and the host. Unlike its oxygen-containing counterpart (5-vinyl-2'-deoxyuridine, which is extremely toxic in vitro and causes chromosome damage), 5-vinyl-4'-thio-2'-deoxyuridine is not toxic and shows a similar antiviral activity spectrum to 5-cyclopropyl- and 5-isopropyl-4'-thio-2'-deoxyuridines which also show no toxicity. Although a good leaving group at the 5'-position of a 4'-thionucleoside appears to be easily displaced (possibly involving intramolecular episulphide formation), when incorporated into an oligodeoxynucleotide, 4'-thiothymidine appears not to cause any gross distortion of the helix as shown by CD or Tm measurements and the phosphodiester backbone is stable to hydrolysis. Thus it is not yet clear whether the toxicity of 4'-thiothymidine is due to its presence in DNA or to the perturbation of the metabolic processes involved in its incorporation.
Assuntos
Oligodesoxirribonucleotídeos/síntese química , Tionucleosídeos/química , Tionucleosídeos/síntese química , Timidina/análogos & derivados , Antivirais/síntese química , Antivirais/química , Sequência de Bases , Citomegalovirus/efeitos dos fármacos , Herpesvirus Humano 3/efeitos dos fármacos , Dados de Sequência Molecular , Núcleosídeo-Fosfato Quinase/química , Oligodesoxirribonucleotídeos/química , Oligodesoxirribonucleotídeos/farmacologia , Fosforilação , Simplexvirus/efeitos dos fármacos , Especificidade por Substrato , Tionucleosídeos/farmacologia , Timidina/químicaRESUMO
We report that a thymidine kinase (TK) activity is present in Trichomonas vaginalis and can be separated from the deoxyribonucleoside phosphotransferase. T. vaginalis TK, purified 11200-fold to apparent homogeneity, has a molecular mass of 31500 Da. It phosphorylates not only thymidine (Km 0.18 microM) but also deoxycytidine (Km 0.88 microM) and deoxyuridine (Km 0.14 microM). In contrast with T. vaginalis deoxyribonucleoside phosphotransferase, the TK activity is strongly inhibited by novel deoxyuridine analogues such as 5-methyl-4'-thio-2'-deoxyuridine (MTdU) (Ki 20 nM) and 5-iodo-4'-thio-2'-deoxyuridine (ITdU) (Ki 24 nM). MTdU and ITdU are phosphorylated by T. vaginalis TK in vitro. In vivo they inhibit [3H]thymidine incorporation in T. vaginalis cultured cells and T. vaginalis growth (IC50 7.5 and 24 microM respectively; minimal lethal dose 100 microM). Thus the TK inhibitors described here demonstrate the key role of T. vaginalis TK for protozoal growth and viability and indicate TK as a new target for the design of antitrichomonal drugs.
Assuntos
Desoxiuridina/farmacologia , Timidina Quinase/metabolismo , Trichomonas vaginalis/enzimologia , Animais , Antiprotozoários/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Cromatografia de Afinidade , Cromatografia Líquida de Alta Pressão , Cromatografia por Troca Iônica , Desoxiuridina/análogos & derivados , Inibidores Enzimáticos/farmacologia , Células HeLa , Humanos , Cinética , Peso Molecular , Especificidade por Substrato , Timidina Quinase/antagonistas & inibidores , Timidina Quinase/química , Timidina Quinase/isolamento & purificação , Trichomonas vaginalis/efeitos dos fármacos , Trichomonas vaginalis/crescimento & desenvolvimentoRESUMO
(E)-5-(2-Bromovinyl)-2'-deoxy-4'-thiouridine (S-BVDU) is a potent antiherpesvirus agent and its use in gene therapy as an anticancer agent has recently been described. We here outline 2 efficient methods for the synthesis of S-BVDU. The decision as to which method is to be used depends upon the starting materials available but starting from BVU, an overall yield of beta-nucleoside of 35% can be expected. From 5-ethyl-2'-deoxy-4'-thiouridine, radical bromination using bromine will give a quantitative conversion to S-BVDU if unreacted starting material is recycled (50%) or using N-bromosuccinimide, a one step yield in excess of 80% can be obtained.
Assuntos
Bromodesoxiuridina/análogos & derivados , Herpes Simples/virologia , Nucleosídeos/síntese química , Tiouridina/análogos & derivados , Antineoplásicos/síntese química , Antivirais/síntese química , Bromodesoxiuridina/síntese química , Bromodesoxiuridina/farmacologia , Estrutura Molecular , Pentosiltransferases/metabolismo , Tiouridina/síntese químicaRESUMO
The synthesis and X-ray crystal structures of a series of 5-substituted-6-aza-2'-deoxyuridines is reported. These nucleoside analogues inhibit the phosphorylation of thymidine by HSV-1 TK but have no effect on the corresponding human enzyme. Detailed examination of one analogue proves it to be a competitive inhibitor of thymidine with a Ki of 0.34 microM and is a very poor substrate. The analogues are not substrates for the enzyme and also do not inhibit the degradation of thymidine by thymidine phosphorylase. Molecular modelling showed that the inhibitors fit well in the active site of HSV-1 TK, provided the conformation of the sugar moiety is the same for thymidine in the complex.
Assuntos
Compostos Aza/síntese química , Azauridina/análogos & derivados , Desoxiuridina/análogos & derivados , Antivirais/síntese química , Compostos Aza/farmacologia , Plaquetas/enzimologia , Cristalografia por Raios X , Inibidores Enzimáticos/síntese química , Herpes Simples/enzimologia , Humanos , Cinética , Espectroscopia de Ressonância Magnética , Modelos Moleculares , Conformação Molecular , Nucleosídeos/síntese química , Nucleosídeos/farmacologia , Fosforilação , Timidina/metabolismo , Timidina Quinase/antagonistas & inibidoresRESUMO
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 4'-thio-2'-deoxyuridines exhibits an anti-[herpes simplex virus (HSV)] activity significantly higher (20-600 times) than that shown by the corresponding 4'-oxy counterpart. We investigated the mode of action of these compounds and we found that: (i) several 4'-thio-2'-deoxyuridines are phosphorylated to the mono- and di-phosphates by HSV-1 thymidine kinase (TK) more efficiently than their corresponding 4'-oxy counterpart; (ii) both are inhibitors of cellular thymidylate synthase; (iii) 4'-thio-2'-deoxyuridines are resistant to phosphorolysis by human thymidine phosphorylase; (iv) both 4'-oxy- and 4'-thio-2'-deoxyuridines are phosphorylated to deoxyribonucleotide triphosphate in HSV-1-infected cells and are incorporated into viral DNA; (v) 4'-thio-2'-deoxyuridines are better inhibitors than their 4'-oxy counterparts of [(3)H]thymidine incorporation in HSV-1-infected cells; (vi) 4'-thio-2'-deoxyuridines are not recognized by HSV-1 and human uracil-DNA glycosylases. Our data suggest that 4'-thio-2'-deoxyuridines, resistant to pyrimidine phosphorylase, can be preferentially or selectively phosphorylated by viral TK in HSV-infected cells, where they are further converted into triphosphate by cellular nucleotide kinases. Once incorporated into viral DNA, they are better inhibitors of viral DNA synthesis than their corresponding 4'-oxy counterpart, either because they are not recognized, and thus not removed, by viral uracil-DNA glycosylase, or because they preferentially interfere with viral DNA polymerase.