RESUMO
Starting from 2-C-methyl-ribonolactone, 1,2,3,5-tetra-O-acetyl-2-C-methyl-4-thioribofuranose was synthesized and condensed with heterocyclic bases to afford 2-C-methyl-4'-thioribonucleosides.
Assuntos
Desoxicitidina/análogos & derivados , Tionucleosídeos/química , Antivirais/farmacologia , Configuração de Carboidratos , Sequência de Carboidratos , Desoxicitidina/farmacologia , Modelos Químicos , Nucleosídeos/química , Vírus de RNA/genética , Tionucleosídeos/síntese químicaRESUMO
In order to evaluate their antiviral properties, two imidazo[4,5-d]pyridazine 2'-C-methyl-beta-D-ribofuranonucleoside derivatives have been synthesized.
Assuntos
Desoxirribonucleosídeos/síntese química , Biologia Molecular/métodos , Nucleosídeos de Purina/química , Piridazinas/química , Antivirais/síntese química , Antivirais/farmacologia , Química Farmacêutica/métodos , Desenho de Fármacos , Modelos Químicos , Piridazinas/síntese química , Ribose/química , Temperatura , Fatores de TempoRESUMO
Coupling suitable sugars (D- or L-ribofuranose, 2' or 3-deoxysugar, branched sugars) with 2-aminoimidazo[1,2-a]-s-triazin-4-one was carried out using the different reaction conditions: 1) condensation in the presence of sodium hydride; or 2) condensation using Vorbrüggen's methods. The 5-aza- 7-deazaguanine nucleoside analogues obtained were evaluated in cell culture experiments for the inhibition of the replication of a number of RNA viruses, including BVDV, YFV, and WNV.
Assuntos
Antivirais/farmacologia , Infecções por Flavivirus/tratamento farmacológico , Flavivirus/metabolismo , Guanina/análogos & derivados , Guanosina/análogos & derivados , Antivirais/química , Química Farmacêutica/métodos , Desenho de Fármacos , Guanina/síntese química , Guanosina/síntese química , Humanos , Modelos Químicos , Nucleosídeos/química , Nucleosídeos de Pirimidina/química , Ribonucleosídeos/químicaRESUMO
In order to evaluate their antiviral properties, a series of 4'-C-methyl-beta-D-ribofuranosyl purine and pyrimidine nucleosides has been prepared. Unfortunately, none of these 4'-branched nucleosides showed any antiviral activity or cytotoxcity when tested against HIV, HBV, and Yellow Fever virus.
Assuntos
Nucleosídeos de Purina/síntese química , Nucleosídeos de Pirimidina/síntese química , Antivirais/síntese química , Desenho de Fármacos , Indicadores e Reagentes , Metilação , Nucleosídeos de Purina/química , Nucleosídeos de Pirimidina/química , RiboseRESUMO
beta-L-Thymidine (L-dT) and beta-L-2'-deoxycytidine (L-dC) are potent and highly specific inhibitors of hepatitis B virus (HBV) replication both in vivo and in vitro (50% effective concentrations, 0.19 to 0.24 microM in 2.2.15 cells). The intracellular metabolisms of L-dT and L-dC were investigated in HepG2 cells and primary cultured human hepatocytes. L-dT and L-dC were extensively phosphorylated in both cell types, with the 5'-triphosphate derivative being the predominant metabolite. In HepG2 cells, the 5'-triphosphate levels were 27.7 +/- 12.1 and 72.4 +/- 1.8 pmol/10(6) cells for L-dT and L-dC, respectively. In primary human hepatocytes, the 5'-triphosphate levels were 16.5 +/- 9.8 and 90.1 +/- 36.4 pmol/10(6) cells for L-dT and L-dC, respectively. Furthermore, a choline derivative of L-dCDP was detected at concentrations of 15.8 +/- 1.8 and 25.6 +/- 0.1 pmol/10(6) cells in human hepatocytes and HepG2 cells, respectively. In HepG2 cells exposed to L-dC, the 5'-monophosphate and 5'-triphosphate derivatives of beta-L-2'-deoxyuridine (L-dUMP and L-dUTP, respectively) were also observed, reaching intracellular concentrations of 6.7 +/- 0.4 and 18.2 +/- 1.0 pmol/10(6) cells, respectively. In human hepatocytes, L-dUMP and L-dUTP were detected at concentrations of 5.7 +/- 2.4 and 43.5 +/- 26.8 pmol/10(6) cells, respectively. It is likely that deamination of L-dCMP by deoxycytidylate deaminase leads to the formation of L-dUMP, as the parent compound, L-dC, was not a substrate for deoxycytidine deaminase. The intracellular half-lives of L-dTTP, L-dCTP, and L-dUTP were at least 15 h, with intracellular concentrations of each metabolite remaining above their respective 50% inhibitory concentrations for the woodchuck hepatitis virus DNA polymerase for as long as 24 h after removal of the drug from cell cultures. Exposure of HepG2 cells to L-dT in combination with L-dC led to concentrations of the activated metabolites similar to those achieved with either agent alone. These results suggest that the potent anti-HBV activities of L-dT and L-dC are associated with their extensive phosphorylation.
Assuntos
Antivirais/farmacologia , Carcinoma Hepatocelular/metabolismo , Desoxicitidina/farmacologia , Vírus da Hepatite B/efeitos dos fármacos , Hepatite B/tratamento farmacológico , Hepatócitos/efeitos dos fármacos , Neoplasias Hepáticas/metabolismo , Timidina/farmacologia , Cromatografia Líquida de Alta Pressão , Desoxicitidina/metabolismo , Meia-Vida , Hepatite B/virologia , Humanos , Fosforilação , Timidina/metabolismo , Células Tumorais CultivadasRESUMO
Three simple, related nucleosides, beta-L-2'-deoxycytidine (LdC), beta-Lthymidine (LdT), and beta-L-2'-deoxyadenosine (LdA), have been discovered to be potent, specific and selective inhibitors of the replication hepatitis B virus (HBV), as well as the closely related duck and woodchuck hepatitis viruses (WHV). Structure-activity relationship analysis indicates that the 3'-OH group of the beta-L-2'-deoxyribose of the beta-L-2'-deoxynucleoside confers specific anti-hepadnavirus activity. The simple nucleosides had no effect on the replication of 15 other RNA and DNA viruses, and did not inhibit human DNA polymerases (alpha, beta and gamma) or compromise mitochondrial function. The nucleosides are efficiently converted intracellularly into active triphosphate metabolites that have a long half-life. Once-daily oral administration of these compounds in the woodchuck efficacy model of chronic HBV infection reduced viral load by as much as 10(8) genome equivalents/ml serum and there was no drug-related toxicity. In addition, a decline in WHV surface antigen (WHsAg) paralleled the decrease in viral load. This class of nucleosides displays an excellent overall safety profile. The first compound, LdT, has already entered clinical trials and LdC, currently being developed as a prodrug, is expected to enter the clinic in the near future. These compounds have the potential for use in combination therapy with the goal of achieving superior viral suppression and diminishing the onset of resistance.
Assuntos
Antivirais/uso terapêutico , Hepatite B/tratamento farmacológico , Nucleosídeos/uso terapêutico , Animais , Antivirais/farmacocinética , Modelos Animais de Doenças , Humanos , Testes de Sensibilidade Microbiana , Nucleosídeos/farmacocinéticaRESUMO
A unique series of simple unnatural L-nucleosides that specifically inhibit hepatitis B virus (HBV) replication has been discovered. These molecules have in common a hydroxyl group in the 3'-position (3'-OH) of the beta-L-2'-deoxyribose sugar that confers antiviral activity specifically against hepadnaviruses. Replacement of the 3'-OH broadens activity to other viruses. Substitution in the base decreases antiviral potency and selectivity. Human DNA polymerases and mitochondrial function are not effected. Plasma viremia is reduced up to 8 logs in a woodchuck model of chronic HBV infection. These investigational drugs, used alone or in combination, are expected to offer new therapeutic options for patients with chronic HBV infection.
Assuntos
Antivirais/farmacologia , Desoxirribonucleosídeos/farmacologia , Vírus da Hepatite B/efeitos dos fármacos , Animais , Antivirais/química , Desoxiadenosinas/química , Desoxiadenosinas/farmacologia , Desoxicitidina/química , Desoxicitidina/farmacologia , Desoxirribonucleosídeos/química , Vírus da Hepatite B da Marmota/efeitos dos fármacos , Vírus da Hepatite B da Marmota/fisiologia , Vírus da Hepatite B/fisiologia , Hepatite B Crônica/tratamento farmacológico , Humanos , Relação Estrutura-Atividade , Especificidade por Substrato , Timidina/química , Timidina/farmacologia , Replicação Viral/efeitos dos fármacosRESUMO
A unique series of simple "unnatural" nucleosides has been discovered to inhibit hepatitis B virus (HBV) replication. Through structure-activity analysis it was found that the 3'-OH group of the beta-L-2'-deoxyribose of the beta-L-2'-deoxynucleoside confers specific antihepadnavirus activity. The unsubstituted nucleosides beta-L-2'-deoxycytidine, beta-L-thymidine, and beta-L-2'-deoxyadenosine had the most potent, selective, and specific antiviral activity against HBV replication. Human DNA polymerases (alpha, beta, and gamma) and mitochondrial function were not affected. In the woodchuck model of chronic HBV infection, viral load was reduced by as much as 10(8) genome equivalents/ml of serum and there was no drug-related toxicity. In addition, the decline in woodchuck hepatitis virus surface antigen paralleled the decrease in viral load. These investigational drugs, used alone or in combination, are expected to offer new therapeutic options for patients with chronic HBV infection.
Assuntos
Antivirais/farmacologia , Vírus da Hepatite B/efeitos dos fármacos , Hepatite B/tratamento farmacológico , Nucleosídeos/farmacologia , Animais , Fármacos Anti-HIV/farmacologia , Antivirais/uso terapêutico , Células da Medula Óssea/efeitos dos fármacos , Linhagem Celular , DNA Viral/biossíntese , DNA Polimerase Dirigida por DNA/metabolismo , Desoxiadenosinas/farmacologia , Desoxiadenosinas/uso terapêutico , Desoxicitidina/farmacologia , Desoxicitidina/uso terapêutico , Feminino , HIV-1/efeitos dos fármacos , Hepatite B/virologia , Humanos , Masculino , Marmota , Nucleosídeos/uso terapêutico , Células-Tronco/efeitos dos fármacos , Timidina/farmacologia , Timidina/uso terapêutico , Replicação Viral/efeitos dos fármacosRESUMO
The syntheses of furanthiofurin [5beta-D-(4'-thioribofuranosyl)furan-3-carboxamide, 1] and thiophenthiofurin [5beta-D-(4'-thioribofuranosyl)thiophene-3-carboxamide, 2], two C-thioribonucleoside analogues of tiazofurin, are described. Direct trifluoroacetic acid-catalyzed C-glycosylation of ethyl furan-3-carboxylate with 1-O-acetyl-2,3,5-tri-O-benzyl-4-thio-D-ribofuranose gave 2- and 5-glycosylated regioisomers, as a mixture of alpha and beta anomers. Ethyl 5-(2,3,5-tri-O-benzyl)-beta-D-(4'-thioribofuranosyl)furan-3-carboxylate (6beta) was debenzylated and then converted into the corresponding amide (furanthiofurin) by reaction with ammonium hydroxide. A similar C-glycosylation of ethyl thiophene-3-carboxylate with 1,2,3,5-tetra-O-acetyl-4-thio-D-ribofuranose catalyzed by stannic chloride afforded an anomeric mixture of 2- and 5-glycosylated regioisomers. Deacetylation of ethyl 5-(2,3,5-tri-O-acetyl)-beta-D-(4'-thioribofuranosyl)thiophene-3-carboxylate (13beta) with methanolic ammonia and treatment of the ethyl ester with ammonium hydroxide gave thiophenthiofurin. The glycosylation site and anomeric configuration were established by (1)H NMR spectroscopy. Thiophenthiofurin was found to be cytotoxic in vitro toward human myelogenous leukemia K562, albeit 39-fold less than thiophenfurin, while furanthiofurin proved to be inactive. K562 cells incubated with thiophenthiofurin resulted in inhibition of inosine 5'-monophosphate dehydrogenase (IMPDH) and an increase in IMP pools with a concurrent decrease in GTP levels. From computational studies it was deduced that, among the C-nucleoside analogues of tiazofurin, activity requires an electrophilic sulfur adjacent to the C-glycosidic bond and an energetically favorable conformer around chi = 0 degrees. Among these, the more constrained (least flexible) compounds (tiazofurin and thiophenfurin) are more active than the less constrained thiophenthiofurin. Those compounds which contain a nucleophilic oxygen in place of the thiazole or thiophene (oxazofurin, furanfurin, and furanthiofurin) show the least activity.