RESUMO
To extend the antiviral properties of 2- and 3-fluoro-3-deazaneplanocins into the evolving 3-deaza-1',6'-isoneplanocin library, 2- (11) and 3-fluoro-1',6'-iso-3-deazaneplanocin A (12) have been explored. The requisite synthesis began with an Ullmann reaction by coupling of a protected cyclopentenyl iodide with either 2-fluoro- or 3-fluoro-3-deazaadenine. Target 12 displayed significant activity towards 5 viruses (µM): H1N1 (EC50 < 0.36, CC50 > 357, SI > 1000), hepatitis B virus (EC50 1.28, CC50 > 357, SI > 279), norovirus (EC50 0.64, CC50 > 357, SI > 558), Ebola (EC50 < 0.1, CC50 > 100, SI > 1000), and Marburg (EC50 < 0.1, CC50 > 100, SI > 1000). On the other hand, while 11 showed limited antiviral effects, its toxicity was significant, precluding any further usefulness.
Assuntos
Ebolavirus , Doença pelo Vírus Ebola , Vírus da Influenza A Subtipo H1N1 , Humanos , Doença pelo Vírus Ebola/tratamento farmacológico , Adenosina , Antivirais/farmacologiaRESUMO
3-deazaneplanocin A (DzNep) and its 3-brominated analogs inhibit replication of several RNA viruses. This antiviral activity is attributed to inhibition of S-adenosyl homocysteine hydrolase (SAHase) and consequently inhibition of viral methyltransferases, impairing translation of viral transcripts. The L-enantiomers of some derivatives retain antiviral activity despite dramatically reduced inhibition of SAHase in vitro. To better understand the mechanisms by which these compounds exert their antiviral effects, we compared DzNep, its 3-bromo-derivative, CL123, and the related enantiomers, CL4033 and CL4053, for their activities towards the model negative-sense RNA virus vesicular stomatitis virus (VSV). In cell culture, DzNep, CL123 and CL4033 each exhibited 50 percent inhibitory concentrations (IC50s) in the nanomolar range whereas the IC50 for the L-form, CL4053, was 34-85 times higher. When a CL123-resistant mutant (VSVR) was selected, it exhibited cross-resistance to each of the neplanocin analogs, but retained sensitivity to the adenosine analog BCX4430, an RNA chain terminator. Sequencing of VSVR identified a mutation in the C-terminal domain (CTD) of the viral large (L) protein, a domain implicated in regulation of L protein methyltransferase activity. CL123 inhibited VSV viral mRNA 5' cap methylation, impaired viral protein synthesis and decreased association of viral mRNAs with polysomes. Modest impacts on viral transcription were also demonstrated. VSVR exhibited partial resistance in each of these assays but its replication was impaired, relative to the parent VSV, in the absence of the inhibitors. These data suggest that DzNep, CL123 and CL4033 inhibit VSV through impairment of viral mRNA cap methylation and that the L-form, CL4053, based on the cross-resistance of VSVR, may act by a similar mechanism.
Assuntos
Adenosina/análogos & derivados , Antivirais/farmacologia , Vírus da Estomatite Vesicular Indiana/efeitos dos fármacos , Replicação Viral/efeitos dos fármacos , Adenosina/química , Adenosina/farmacologia , Animais , Chlorocebus aethiops , Concentração Inibidora 50 , Metilação/efeitos dos fármacos , Biossíntese de Proteínas/efeitos dos fármacos , Inibidores da Síntese de Proteínas/farmacologia , Transcrição Gênica/efeitos dos fármacos , Células Vero , Vírus da Estomatite Vesicular Indiana/genéticaRESUMO
In seeking to increase the library of fluorine containing adenine-derived carbocyclic nucleoside antiviral candidates, d-like and l-like 6'-fluoro-3-deazaneplanocin and its 3-bromo derivative lacking the 4'-hydroxylmethylene substituent (2/3 and 4/5, respectively) are presented. Their synthesis was accomplished from d-ribose by developing a more facile precursor route than suggested by the literature. The 2/4d-like pair displayed significant anti-filo virial properties while the enantiomeric l-like congeners 3/5 were inactive. Target compounds 2/4 also were active towards measles and norovirus. The effect of 2/4 is further evidence of the role fluoro-derived adenine carbocyclic nucleoside can play in antiviral drug discovery. Furthermore, the simplicity of their synthesis lends them to more efficacious analogs and to scale-up optimization. There were no other relevant antiviral properties for 2/3 and 4/5 (except BK polyomavirus for 3/5).
Assuntos
Adenosina/análogos & derivados , Antivirais/farmacologia , Ebolavirus/efeitos dos fármacos , Marburgvirus/efeitos dos fármacos , Adenosina/síntese química , Adenosina/química , Adenosina/farmacologia , Antivirais/síntese química , Antivirais/química , Relação Dose-Resposta a Droga , Estrutura Molecular , Estereoisomerismo , Relação Estrutura-AtividadeRESUMO
The arbocyclic nucleosides aristeromycin and neplanocin have been studied as a source for new antiviral agents. A convenient synthesis of C-5'-truncated 3-deaza-1',6'-isoneplanocin, which combines the features of antiviral candidates 5'-noraristeromycin and 3-deaza-1',6'-isoneplanocin is reported from (-)-cyclopentenone to give the two C-4' epimers of 5'-nor-3-deaza isoneplanocin. Antiviral assays showed activity against the JC virus (EC50 = 1.12 µM for (4'R)-8; EC50 = 59.14 µM for (4'S)-7) and inactivity of both compounds against several DNA and RNA viruses. Both compounds lacked cytotoxicity.
Assuntos
Adenosina , Antivirais , Vírus JC/crescimento & desenvolvimento , Vírus de RNA/crescimento & desenvolvimento , Adenosina/análogos & derivados , Adenosina/síntese química , Adenosina/química , Adenosina/farmacologia , Antivirais/síntese química , Antivirais/química , Antivirais/farmacologia , HumanosRESUMO
Enantiomeric 3-deaza-1',6'-isoneplanocins (C-3 unsubstituted 7a/7b and C-3 with a bromine 8a/8b) lacking the 4'-hydroxymethyl as mechanistically designed anti-viral targets have been prepared by utilizing the Ullmann reaction. Anti-Ebola properties were found for the D-like 7a and 8a and L-like 8b. All four products showed effects against human cytomegalovirus while D-like 7a/8a affected measles; 7a was effective versus norovirus and 8a inhibited Pichinde. Both 7a and 8a produced SAHase inhibitory effects. However, the anti-EBOV activity of 7a and 8a cannot be readily correlated with this observation due with their contrasting IC50 values (8aâ¯>â¯7a). It is to be noted that 7b showed no effects on this enzyme and 8b was minimally inhibitory. These results offer preliminary insight into the differing mechanisms of action of D- and L- like structures and enlighten structural features to guide additional antiviral agent pursuit in the isoneplanocin series.
Assuntos
Adenosina/análogos & derivados , Antivirais/síntese química , Adenosina/síntese química , Adenosina/farmacologia , Adenosil-Homocisteinase/antagonistas & inibidores , Adenosil-Homocisteinase/metabolismo , Animais , Antivirais/química , Antivirais/farmacologia , Citomegalovirus/efeitos dos fármacos , Ebolavirus/efeitos dos fármacos , Eritrócitos/enzimologia , Humanos , Norovirus/efeitos dos fármacos , Coelhos , EstereoisomerismoRESUMO
A convenient stereospecific synthesis of 6'-fluoro-3-deazaneplanocin (6) has been accomplished from d-ribose in 15 steps. It is reported to possess significant activity towards Ebola (Zaire, Vero, µM: EC50â¯<â¯0.36; CC50 125; SIâ¯>â¯347) with moderate inhibition of the target enzyme (S-adenosylhomocysteine hydrolase), which did not correlate directly with its anti-Ebola effects. Compound 6, with limited cytotoxicity, also displayed activity against measles, H1N1 and Pichinde.
Assuntos
Adenosina/análogos & derivados , Antivirais/síntese química , Adenosina/síntese química , Adenosina/química , Adenosina/farmacologia , Adenosil-Homocisteinase/antagonistas & inibidores , Adenosil-Homocisteinase/metabolismo , Animais , Antivirais/química , Antivirais/farmacologia , Chlorocebus aethiops , Ebolavirus/efeitos dos fármacos , Vírus da Influenza A Subtipo H1N1/efeitos dos fármacos , Concentração Inibidora 50 , Células VeroRESUMO
Synthetically combining the C-4' side-chain structural features of the antiviral candidates 5'-methylaristeromycin and 5'-homoaristeromycin into a diastereomeric pair of C-4' side-chain dihydroxylated aristeromycins (6 and 7) is reported. Broad antiviral analyses of the both targets found promising effects towards HBV (6, 6.7⯵M and 7, 7.74⯵M) and HCMV (only 7, 0.72⯵M). No other activity was found. Neither of the diastereomers was cytotoxic in the assays performed.
Assuntos
Adenosina/análogos & derivados , Antivirais/farmacologia , Citomegalovirus/efeitos dos fármacos , Vírus da Hepatite B/efeitos dos fármacos , Adenosina/síntese química , Adenosina/química , Adenosina/farmacologia , Antivirais/síntese química , Antivirais/química , Relação Dose-Resposta a Droga , Humanos , Testes de Sensibilidade Microbiana , Conformação Molecular , Relação Estrutura-AtividadeRESUMO
Both enantiomers of the isomer of neplanocin where the C-4' hydroxymethyl has been displaced to the 6'-position (that is, 6'-isoneplanocin) have been prepared in 5 steps from known, protected iodocyclopentenones. Both products were evaluated against a number of DNA and RNA viruses and found to be inactive. This observation is suggested to be due to the displacement of the C-4' hydroxymethyl of neplanocin (in the D-like form) away from the lysine425 of S-adenosylhomocysteine hydrolase, which is important for inhibition by neplanocin and, in turn, its antiviral activity.
Assuntos
Adenosina/análogos & derivados , Antivirais/química , Adenosina/química , Adenosina/farmacologia , Antivirais/farmacologia , Modelos Moleculares , Conformação Molecular , EstereoisomerismoRESUMO
Objective To synthesize 3,7-dideazaneplanocin and evaluate its antiviral potential. Methods The target 3,7-dideazaneplanocin has been prepared in five steps from a readily available cyclopentenol. A thorough in vitro antiviral analysis was conducted versus both DNA and RNA viruses. Results A rational synthesis of 3,7-dideazaneplanocin was conceived and successfully pursued in such a way that it can be adapted to various analogs of 3,7-dideazaneplanocin. Using standard antiviral assays, no activity for 3,7-dideazaneplanocn was found. Conclusion Two structural features are necessary for adenine-based carbocyclic nucleosides (like neplanocin) for potential antiviral properties: (i) inhibition of S-adenosylhomocysteine hydrolase and/or (ii) C-5' activation via the mono-nucleotide. These two requisite adenine structural features to fit these criteria are not present in in the target 3,7-dideazaneplanocin: (i) an N-7 is necessary for inhibition of the hydrolase and the N-3 is claimed to be essential for phosphorylation at C-5'. Thus, it is not surprising that 3,7-dideazaneplaoncin lacked antiviral properties.
Assuntos
Antivirais/farmacologia , Vírus de DNA/efeitos dos fármacos , Inibidores Enzimáticos/farmacologia , Piridonas/farmacologia , Vírus de RNA/efeitos dos fármacos , Antivirais/síntese química , Antivirais/química , Inibidores Enzimáticos/síntese química , Inibidores Enzimáticos/química , Hidrolases/antagonistas & inibidores , Hidrolases/metabolismo , Testes de Sensibilidade Microbiana , Conformação Molecular , Piridonas/síntese química , Piridonas/químicaRESUMO
The 1',6'-isomer of neplanocin A possesses biological properties that have not been optimised through rationally conceived analogues. In that direction, this Letter reports the use of the Ullmann reaction to achieve enantiomeric 3-deaza-1',6'-isoneplanocin and 3-bromo-3-deaza-1',6'-isoneplanocin. These four compounds showed significant Ebola activity that is not specifically due to their inhibition of S-adenonosylhomocysteine hydrolase, as might have been expected for 3-deazaadenine carbocyclic nucleosides. For some members of this group, antiviral activity was also found against human cytomegalovirus, hepatitis B, norovirus, and measles.
Assuntos
Adenosina/análogos & derivados , Antivirais/química , Adenina/análogos & derivados , Adenina/química , Adenosina/síntese química , Adenosina/química , Adenosina/farmacologia , Adenosil-Homocisteinase/antagonistas & inibidores , Adenosil-Homocisteinase/metabolismo , Antivirais/síntese química , Antivirais/farmacologia , Citomegalovirus/efeitos dos fármacos , Ebolavirus/efeitos dos fármacos , Humanos , Morbillivirus/efeitos dos fármacos , Norovirus/efeitos dos fármacos , Nucleosídeos/química , EstereoisomerismoRESUMO
The preparation of the C-5' diastereomers of 5'-homoaristeromycin has been achieved in 6 steps beginning with readily available (3aR,6aR)-2,2-dimethyl-2H,3aH,4H,6aH-cyclopenta[1,3]dioxol-4-one in a Michael reaction employing chiral Evans N-acyloxazolidinones that served to direct the requisite side chain stereochemistry. The two targets were evaluated against a battery of viruses and found to possess activity only towards yellow fever. Both compounds were non-cytotoxic.
Assuntos
Adenosina/análogos & derivados , Antivirais/síntese química , Adenosina/síntese química , Adenosina/química , Adenosina/farmacologia , Antivirais/química , Antivirais/farmacologia , Humanos , Metilação , Oxazolidinonas/síntese química , Oxazolidinonas/química , Estereoisomerismo , Titânio/química , Viroses/tratamento farmacológico , Vírus/efeitos dos fármacosRESUMO
The 3-deaza analogs of the naturally occurring adenine-based carbocyclic nucleosides aristeromycin and neplanocin possess biological properties that have not been optimized. In that direction, this paper reports the strategic placement of a fluorine atom at the C-2 and C-3 positions and a methyl at the C-3 site of the 3-deazaadenine ring of the aforementioned compounds. The synthesis and S-adenosylhomocysteine hydrolase inhibitory and antiviral properties of these targets are described. Some, but not all, compounds in this series showed significant activity toward herpes, arena, bunya, flavi, and orthomyxoviruses.
Assuntos
Adenosina/análogos & derivados , Antivirais/farmacologia , Adenosina/síntese química , Humanos , Estrutura MolecularRESUMO
The naturally occurring adenine based carbocyclic nucleosides aristeromycin and neplanocin A and their 3-deaza analogues have found a prominent place in the search for diverse antiviral activity agent scaffolds because of their ability to inhibit S-adenosylhomocysteine (AdoHcy) hydrolase. Following the lead of these compounds, their 3-deaza-3-fluoroaristeromycin analogues have been synthesized and their effect on S-adenosylhomocysteine hydrolase and RNA and DNA viruses determined.
Assuntos
Antivirais/síntese química , S-Adenosil-Homocisteína/análogos & derivados , Adenosil-Homocisteinase/antagonistas & inibidores , Adenosil-Homocisteinase/metabolismo , Animais , Antivirais/química , Antivirais/farmacologia , Linhagem Celular , Chlorocebus aethiops , Vírus de DNA/efeitos dos fármacos , Inibidores Enzimáticos/síntese química , Inibidores Enzimáticos/química , Células HEK293 , Humanos , Vírus de RNA/efeitos dos fármacos , S-Adenosil-Homocisteína/síntese química , S-Adenosil-Homocisteína/farmacologia , Células VeroRESUMO
To extend the potential of 5'-noraristeromycin (and its enantiomer) as potential antiviral candidates, the enantiomers of the carbocyclic 5'-nor derivatives of 5'-methylthio-5'-deoxyadenosine and 5'-phenylthio-5'-deoxyadenosine have been synthesized and evaluated. None of the compounds showed meaningful antiviral activity.
Assuntos
Antivirais/farmacologia , Desenho de Fármacos , Vírus/efeitos dos fármacos , Adenosina/análogos & derivados , Adenosina/síntese química , Adenosina/química , Adenosina/farmacologia , Adenosil-Homocisteinase/antagonistas & inibidores , Desoxiadenosinas/síntese química , Desoxiadenosinas/química , Desoxiadenosinas/farmacologia , Relação Estrutura-Atividade , Tionucleosídeos/síntese química , Tionucleosídeos/química , Tionucleosídeos/farmacologiaRESUMO
Both enantiomers of 1',6'-isoneplanocin have been prepared from a common substituted cyclopentane epoxide in 7 steps. Both compounds were subjected to DNA and RNA viral assessments with moderate to high activity found for both towards human cytomegalovirus, measles, Ebola, norovirus, and dengue. The D-like congener also showed vaccinia and HBV effectiveness. In many of the other antiviral assays both compounds showed cytotoxicity making, in some cases, an EC50 determination not possible. The S-adenosylhomocysteine hydrolase inhibitory effects showed the D-like target to be equal that of neplanocin itself and better than 3-deazaneplanocin whereas the L-like analogue was 13 to 30 times less inhibitory than 3-deazaneplanocin and neplanocin, respectively.
Assuntos
Adenosina/análogos & derivados , Antivirais/síntese química , Antivirais/farmacologia , Adenosina/síntese química , Adenosina/química , Adenosina/farmacologia , Antivirais/química , Citomegalovirus/efeitos dos fármacos , Vírus da Dengue/efeitos dos fármacos , Relação Dose-Resposta a Droga , Ebolavirus/efeitos dos fármacos , Vírus da Hepatite B/efeitos dos fármacos , Vírus do Sarampo/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Estrutura Molecular , Norovirus/efeitos dos fármacos , Estereoisomerismo , Relação Estrutura-Atividade , Vaccinia virus/efeitos dos fármacosRESUMO
To identify potential antivirals against BTV, we have developed, optimized and validated three assays presented here. The CPE-based assay was the first assay developed to evaluate whether a compound showed any antiviral efficacy and have been used to screen large compound library. Meanwhile, cytotoxicity of antivirals could also be evaluated using the CPE-based assay. The dose-response assay was designed to determine the range of efficacy for the selected antiviral, i.e. 50% inhibitory concentration (IC50) or effective concentration (EC50), as well as its range of cytotoxicity (CC50). The ToA assay was employed for the initial MoA study to determine the underlying mechanism of the novel antivirals during BTV viral lifecycle or the possible effect on host cellular machinery. These assays are vital for the evaluation of antiviral efficacy in cell culture system, and have been used for our recent researches leading to the identification of a number of novel antivirals against BTV.
Assuntos
Antirretrovirais/farmacologia , Vírus Bluetongue/efeitos dos fármacos , Animais , Bluetongue/tratamento farmacológico , Bluetongue/virologia , Linhagem Celular , Cricetinae , Efeito Citopatogênico Viral/efeitos dos fármacos , Relação Dose-Resposta a DrogaRESUMO
BACKGROUND: Tacaribe virus (TCRV) is a less biohazardous relative of the highly pathogenic clade B New World arenaviruses that cause viral hemorrhagic fever syndromes and require handling in maximum containment facilities not readily available to most researchers. AG129 type I and II interferon receptor knockout mice have been shown to be susceptible to TCRV infection, but the pathogenic mechanisms contributing to the lethal disease are unclear. METHODS: To gain insights into the pathogenesis of TCRV infection in AG129 mice, we assessed hematologic and cytokine responses during the course of infection, as well as changes in the permeability of the vascular endothelium. We also treated TCRV-challenged mice with MY-24, a compound that prevents mortality without affecting viral loads during the acute infection, and measured serum and tissue viral titers out to 40 days post-infection to determine whether the virus is ultimately cleared in recovering mice. RESULTS: We found that the development of viremia and splenomegaly precedes an elevation in white blood cells and the detection of high levels of proinflammatory mediators known to destabilize the endothelial barrier, which likely contributes to the increased vascular permeability and weight loss that was observed several days prior to when the mice generally succumb to TCRV challenge. In surviving mice treated with MY-24, viremia and liver virus titers were not cleared until 2-3 weeks post-infection, after which the mice began to recover lost weight. Remarkably, substantial viral loads were still present in the lung, spleen, brain and kidney tissues at the conclusion of the study. CONCLUSIONS: Our findings suggest that vascular leak may be a contributing factor in the demise of TCRV-infected mice, as histopathologic findings are generally mild to moderate in nature, and as evidenced with MY-24 treatment, animals can survive in the face of high viral loads.
Assuntos
Infecções por Arenaviridae/imunologia , Infecções por Arenaviridae/patologia , Arenavirus do Novo Mundo/imunologia , Arenavirus do Novo Mundo/patogenicidade , Permeabilidade Capilar , Citocinas/metabolismo , Estruturas Animais/virologia , Animais , Modelos Animais de Doenças , Feminino , Humanos , Masculino , Camundongos , Camundongos Knockout , Carga Viral , Viremia/imunologia , Viremia/patologiaRESUMO
The synthesis of 6'-methyl-5'-homoaristeromycin is described from a known 6'-ethyl ester. Antiviral analysis showed the (S)-6' stereoisomer retained the vaccinia activity of the parent 5'-homoaristeromycin (1) while the (R)-6' isomer was less active. Both were weaker than 1 towards cowpox. The diastereomers were equally active versus Epstein Barr virus while (S)-6' was three times more active toward vesicular stomatitis virus than (R)-6'. The diastereomers were inactive towards numerous other viruses. The CC50 for both diastereomers was >300µM.
Assuntos
Adenosina/análogos & derivados , Orthopoxvirus/efeitos dos fármacos , Adenosina/síntese química , Adenosina/química , Adenosina/farmacologia , Animais , Antivirais/síntese química , Antivirais/química , Antivirais/farmacologia , Humanos , Isomerismo , Estrutura MolecularRESUMO
To expand on the antiviral properties of 5'-noraristeromycin, synthetic entry into 3-substituted 3-deaza-5'-noraristeromyin derivatives (i.e., bromo, 4; iodo, 5; chloro, 6; and, methyl, 7) has been accomplished from a common intermediate. An extensive antiviral analysis showed 7 to be basically inactive (except for weak effects against VSV) and there were no general trends among the halo compounds (except versus reovirus-1 and influenza B). Individually, compound 4 was most favorable towards HCMV, VZV, HBV, and VV; product 5 against HBV, VSV, VV, influenza B, HCMV, and measles; and, target 6 towards Punta Toro, VSV, measles, parainflucenza-3, influenza A (H5N1), and influenza B. The methyl target 7 was inactive in all viral assays.
Assuntos
Adenosina/análogos & derivados , Antivirais/química , Antivirais/farmacologia , Vírus/efeitos dos fármacos , Adenina/análogos & derivados , Adenina/síntese química , Adenina/química , Adenina/farmacologia , Adenosina/síntese química , Adenosina/química , Adenosina/farmacologia , Antivirais/síntese química , Cristalografia por Raios X , Humanos , Modelos Moleculares , Viroses/tratamento farmacológicoRESUMO
Bluetongue virus (BTV), a member in the family Reoviridae, is a re-emerging animal disease infecting cattle and sheep. With its recent outbreaks in Europe, there is a pressing need for efficacious antivirals. We presented here the identification and characterization of a novel virostatic molecule against BTV, an aminothiophenecarboxylic acid derivative named compound 003 (C003). The virostatic efficacy of C003 could be improved via chemical modification, leading to a de novo synthesized compound 052 (C052). The 50% effective concentrations (EC(50)) of C003 and C052 were determined at 1.76 ± 0.73 µM and 0.27 ± 0.12 µM, respectively. The 50% cytotoxicity concentration (CC(50)) of C003 was over 100 µM and the CC(50) of C052 was at 82.69 µM. Accordingly, the 50% selective index (SI(50)) of C003 and C052 against BTV was over 57 and 306, respectively. The inhibitory effect of C003/C052 on BTV-induced apoptosis was also confirmed via the inhibition of caspase-3/-7 activation post BTV infection. C003/C052 could inhibit BTV induced CPE even when added as late as 24 h.p.i., indicating that they might act at late stage of viral life-cycle. C003/C052 could reduce over two-logs of both the progeny virus production and the number of genomic viral RNA copies. Interestingly, both the activation of host autophagy and viral protein expression were inhibited post BTV infection when cells were treated with C003 and C052, suggesting that C003/C052 might act as virostatic agents via inhibiting host autophagy activation. Although further investigations might be needed to pin down the exact mechanism of C003/C052, our finding suggested that these compounds might be potent lead compounds with potential novel mechanism of action against BTV.