RESUMEN
Lipid A is an essential component of the Gram negative outer membrane, which protects the bacterium from attack of many antibiotics. The Lipid A biosynthesis pathway is essential for Gram negative bacterial growth and is unique to these bacteria. The first committed step in Lipid A biosynthesis is catalysis by LpxC, a zinc dependent deacetylase. We show the design of an LpxC inhibitor utilizing a robust model which directed efficient design of picomolar inhibitors. Analysis of physiochemical properties drove design to focus on an optimal lipophilicity profile. Further structure based design took advantage of a conserved water network over the active site, and with the optimal lipophilicity profile, led to an improved LpxC inhibitor with in vivo activity against wild type Pseudomonas aeruginosa.
Asunto(s)
Amidohidrolasas/química , Antibacterianos/síntesis química , Inhibidores Enzimáticos/síntesis química , Ácidos Hidroxámicos/síntesis química , Pseudomonas aeruginosa/efectos de los fármacos , Amidohidrolasas/antagonistas & inhibidores , Antibacterianos/farmacología , Dominio Catalítico , Diseño de Fármacos , Inhibidores Enzimáticos/farmacología , Interacciones Hidrofóbicas e Hidrofílicas , Ácidos Hidroxámicos/farmacología , Lípido A/metabolismo , Pruebas de Sensibilidad Microbiana , Modelos Moleculares , Unión Proteica , Pseudomonas aeruginosa/enzimología , Relación Estructura-Actividad , Agua/químicaRESUMEN
A versatile synthesis of 4-oxo-4,7-dihydrofuro[2,3-b]pyridine-5-carboxylate esters has been developed which has lead to the identification of a new series of non-nucleoside inhibitors of human herpesvirus polymerases HCMV, HSV-1, EBV, and VZV with high specificity compared to human DNA polymerases.
Asunto(s)
Antivirales/síntesis química , Inhibidores Enzimáticos/síntesis química , Furanos/síntesis química , Herpesviridae/efectos de los fármacos , Inhibidores de la Síntesis del Ácido Nucleico , Piridonas/síntesis química , Antivirales/farmacología , Química Farmacéutica/métodos , Citomegalovirus/enzimología , Diseño de Fármacos , Inhibidores Enzimáticos/farmacología , Ésteres , Furanos/farmacología , Herpesvirus Humano 3/enzimología , Herpesvirus Humano 4/enzimología , Concentración 50 Inhibidora , Modelos Químicos , Conformación Molecular , Piridonas/farmacología , Simplexvirus/enzimologíaRESUMEN
Oxazolidinones possessing a C-5 carboxamide functionality (reverse amides) represent a new series of compounds that block bacterial protein synthesis. These reverse amides also exhibited less potency against monoamine oxidase (MAO) enzymes and thus possess less potential for the side effects associated with MAO inhibition. The title compound (14) showed reduced in vivo myelotoxicity compared to linezolid in a 14-day safety study in rats, potent in vivo efficacy in murine systemic infection models, and excellent pharmacokinetic properties.
Asunto(s)
Antibacterianos/síntesis química , Óxidos S-Cíclicos/síntesis química , Oxazolidinonas/síntesis química , Acetamidas/farmacología , Administración Oral , Animales , Antibacterianos/farmacología , Antibacterianos/toxicidad , Disponibilidad Biológica , Óxidos S-Cíclicos/farmacología , Óxidos S-Cíclicos/toxicidad , Perros , Farmacorresistencia Bacteriana , Femenino , Bacterias Gramnegativas/efectos de los fármacos , Bacterias Grampositivas/efectos de los fármacos , Inyecciones Intravenosas , Linezolid , Masculino , Ratones , Pruebas de Sensibilidad Microbiana , Inhibidores de la Monoaminooxidasa/síntesis química , Inhibidores de la Monoaminooxidasa/farmacología , Inhibidores de la Monoaminooxidasa/toxicidad , Oxazolidinonas/farmacología , Oxazolidinonas/toxicidad , Ratas , Ratas Sprague-Dawley , Infecciones Estafilocócicas/tratamiento farmacológico , Staphylococcus aureus , Infecciones Estreptocócicas/tratamiento farmacológico , Streptococcus pyogenes , Relación Estructura-ActividadRESUMEN
A novel series of 4-oxo-4,7-dihydrothieno[2,3-b]pyridine-5-carboxamides have been identified as potential antivirals against human herpesvirus infections resulting from human cytomegalovirus (HCMV), herpes simplex virus type 1 (HSV-1), and varicella-zoster virus (VZV). Compounds 10c and 14 demonstrated broad-spectrum inhibition of the herpesvirus polymerases HCMV, HSV-1, and VZV. High specificity for the viral polymerases was observed compared to human alpha polymerase. The antiviral activity of 10c and 14, as determined by plaque reduction assay, was comparable or superior to that of existing antiherpes drugs, ganciclovir (for HCMV) and acyclovir (for HSV-1 and VZV). Drug resistance to compound 14 correlated to point mutations in conserved domain III of the herpesvirus DNA polymerase, but these mutations do not confer resistance to existing nucleoside therapy. In addition, compound 14 maintained potent antiviral activity against acyclovir-resistant HSV-1 strains. Substitution to the pyridone nitrogen (N7) was found to be critical for enhanced in vitro antiviral activity.
Asunto(s)
Antivirales/síntesis química , Citomegalovirus/efectos de los fármacos , Herpesvirus Humano 1/efectos de los fármacos , Herpesvirus Humano 3/efectos de los fármacos , Inhibidores de la Síntesis del Ácido Nucleico , Piridinas/síntesis química , Piridonas/síntesis química , Tiofenos/síntesis química , Animales , Antivirales/química , Antivirales/farmacología , Línea Celular , Supervivencia Celular , Chlorocebus aethiops , Citomegalovirus/enzimología , ADN Polimerasa Dirigida por ADN/química , ADN Polimerasa Dirigida por ADN/genética , Farmacorresistencia Viral , Exodesoxirribonucleasas/antagonistas & inhibidores , Exodesoxirribonucleasas/genética , Herpesvirus Humano 1/enzimología , Herpesvirus Humano 3/enzimología , Humanos , Mutación Puntual , Piridinas/química , Piridinas/farmacología , Piridonas/química , Piridonas/farmacología , Relación Estructura-Actividad , Tiofenos/química , Tiofenos/farmacología , Ensayo de Placa Viral , Proteínas Virales/antagonistas & inhibidores , Proteínas Virales/genéticaRESUMEN
Herpesviruses cause a wide variety of human diseases ranging from cold sores and genital herpes to encephalitis, congenital infections and lymphoproliferative diseases. These opportunistic viruses cause major problems in immunocompromised individuals such as transplant recipients, cancer patients, and HIV-infected persons. The current treatment of these infections is not optimal and there is a need for more active, less toxic compounds that might be used in place of or in addition to current therapies. We have evaluated a new series of 4-oxo-dihydroquinolines, which have a different mechanism of action than nucleosides and have activity against multiple herpesviruses. Of the four new compounds evaluated, two (PHA-529311 and PHA-570886) had greater activity than the parent, PHA-183792, against several herpesviruses and one (PHA-568561) was as effective as the parent. A fourth, PHA-243672, was considerably less effective. They had greater efficacy against cytomegalovirus (CMV) than the other herpesviruses tested and also had activity against acyclovir-resistant herpes simplex virus and varicella-zoster virus isolates and ganciclovir or foscarnet-resistant CMV isolates. These results confirm the broad-spectrum efficacy of these compounds against multiple herpesviruses and suggest that members of this class may have a potential role for treatment of a variety of herpesvirus infections.
Asunto(s)
Antivirales/farmacología , Herpesviridae/efectos de los fármacos , Quinolinas/farmacología , Antivirales/química , Células Cultivadas , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/farmacología , Herpesviridae/enzimología , Herpesviridae/fisiología , Humanos , Pruebas de Sensibilidad Microbiana , Inhibidores de la Síntesis del Ácido Nucleico , Quinolinas/química , Ensayo de Placa Viral , Replicación Viral/efectos de los fármacosRESUMEN
We identified a novel class of 4-oxo-dihydroquinolines represented by PNU-183792 which specifically inhibit herpesvirus polymerases. PNU-183792 was highly active against human cytomegalovirus (HCMV, IC(50) value 0.69 microM), varicella zoster virus (VZV, IC(50) value 0.37 microM) and herpes simplex virus (HSV, IC(50) value 0.58 microM) polymerases but was inactive (IC(50) value >40 microM) against human alpha (alpha), gamma (gamma), or delta (delta) polymerases. In vitro antiviral activity against HCMV was determined using cytopathic effect, plaque reduction and virus yield reduction assays (IC(50) ranging from 0.3 to 2.4 microM). PNU-183792 antiviral activity against both VZV (IC(50) value 0.1 microM) and HSV (IC(50) ranging from 3 to 5 microM) was analyzed using plaque reduction assays. PNU-183792 was also active (IC(50) ranging 0.1-0.7 microM) in cell culture assays against simian varicella virus (SVV), murine cytomegalovirus (MCMV) and rat cytomegalovirus (RCMV). Cell culture activity was compared with the appropriate licensed drugs ganciclovir (GCV), cidofovir (CDV) and acyclovir (ACV). PNU-183792 was also active against both GCV-resistant and CDV-resistant HCMV and against ACV-resistant HSV. Toxicity assays using four different species of proliferating mammalian cells indicated PNU-183792 was not cytotoxic at relevant drug concentrations (CC(50) value >100 microM). PNU-183792 was inactive against unrelated DNA and RNA viruses indicating specificity for herpesviruses. In animals, PNU-183792 was orally bioavailable and was efficacious in a model of lethal MCMV infection.
Asunto(s)
Antivirales/farmacología , Inhibidores Enzimáticos/farmacología , Infecciones por Herpesviridae/tratamiento farmacológico , Herpesviridae/efectos de los fármacos , Inhibidores de la Síntesis del Ácido Nucleico , Quinolinas/farmacología , Animales , Antivirales/química , Antivirales/farmacocinética , Línea Celular , Farmacorresistencia Viral , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/farmacocinética , Herpesviridae/enzimología , Infecciones por Herpesviridae/virología , Humanos , Ratones , Ratones Endogámicos BALB C , Muromegalovirus/efectos de los fármacos , Quinolinas/química , Quinolinas/farmacocinética , Ratas , Ratas Sprague-DawleyRESUMEN
We report a new class of non-nucleoside antivirals, the 7-oxo-4,7-dihydrothieno[3,2-b]pyridine-6-carboxamides, some of which possess remarkable potency versus a broad spectrum of herpesvirus DNA polymerases and excellent selectivity compared to human DNA polymerases. A critical factor in the level of activity is hypothesized to be conformational restriction of the key 2-aryl-2-hydroxyethylamine sidechain by an adjacent methyl group.
Asunto(s)
Citomegalovirus/enzimología , Inhibidores Enzimáticos/síntesis química , Inhibidores Enzimáticos/farmacología , Inhibidores de la Síntesis del Ácido Nucleico , Piridinas/química , Piridinas/síntesis química , Piridinas/farmacología , Relación Estructura-ActividadRESUMEN
A novel series of 2-aryl-2-hydroxyethylamine substituted 4-oxo-4,7-dihydrothieno[2,3-b]pyridine-5-carboxamides have been identified as potent antivirals against human herpesviruses. These compounds demonstrate broad-spectrum inhibition of the herpesvirus polymerases HCMV, HSV-1, EBV, and VZV with high specificity compared to human DNA polymerases.
Asunto(s)
Antivirales/farmacología , Citomegalovirus/efectos de los fármacos , Inhibidores Enzimáticos/farmacología , Etilaminas/farmacología , Exodesoxirribonucleasas/antagonistas & inhibidores , Herpesviridae/efectos de los fármacos , Herpesvirus Humano 1/efectos de los fármacos , Herpesvirus Humano 4/efectos de los fármacos , Inhibidores de la Síntesis del Ácido Nucleico , Piridinas/farmacología , Proteínas Virales/antagonistas & inhibidores , Antivirales/síntesis química , Antivirales/uso terapéutico , Unión Competitiva , ADN Polimerasa Dirigida por ADN , Inhibidores Enzimáticos/síntesis química , Inhibidores Enzimáticos/uso terapéutico , Etilaminas/síntesis química , Herpes Zóster/tratamiento farmacológico , Herpesviridae/enzimología , Humanos , Modelos Químicos , Piridinas/síntesis química , Relación Estructura-ActividadRESUMEN
The susceptibility of human cytomegalovirus (CMV) and varicella zoster virus (VZV) clinical isolates to PNU-183792, a 4-oxo-dihydroquinoline, was examined. The antiviral potency of PNU-183792, a non-nucleoside inhibitor, was compared to the licensed nucleoside inhibitors ganciclovir and acyclovir using plaque reduction and virus yield reduction assays. PNU-183792 was as potent against CMV as ganciclovir and was superior in potency to acyclovir against VZV. PNU-183792 represents a new class of non-nucleoside inhibitors of human herpesviruses.
Asunto(s)
Antivirales/farmacología , Citomegalovirus/efectos de los fármacos , Herpesvirus Humano 3/efectos de los fármacos , Quinolinas/farmacología , Aciclovir/farmacología , Línea Celular , Citomegalovirus/crecimiento & desarrollo , Citomegalovirus/aislamiento & purificación , Citomegalovirus/fisiología , Ganciclovir/farmacología , Herpesvirus Humano 3/crecimiento & desarrollo , Herpesvirus Humano 3/aislamiento & purificación , Herpesvirus Humano 3/fisiología , Humanos , Técnicas In Vitro , Ensayo de Placa Viral , Replicación Viral/efectos de los fármacosRESUMEN
The 4-oxo-dihydroquinolines (PNU-182171 and PNU-183792) are nonnucleoside inhibitors of herpesvirus polymerases (R. J. Brideau et al., Antiviral Res. 54:19-28, 2002; N. L. Oien et al., Antimicrob. Agents Chemother. 46:724-730, 2002). In cell culture these compounds inhibit herpes simplex virus type 1 (HSV-1), HSV-2, human cytomegalovirus (HCMV), varicella-zoster virus (VZV), and human herpesvirus 8 (HHV-8) replication. HSV-1 and HSV-2 mutants resistant to these drugs were isolated and the resistance mutation was mapped to the DNA polymerase gene. Drug resistance correlated with a point mutation in conserved domain III that resulted in a V823A change in the HSV-1 or the equivalent amino acid in the HSV-2 DNA polymerase. Resistance of HCMV was also found to correlate with amino acid changes in conserved domain III (V823A+V824L). V823 is conserved in the DNA polymerases of six (HSV-1, HSV-2, HCMV, VZV, Epstein-Barr virus, and HHV-8) of the eight human herpesviruses; the HHV-6 and HHV-7 polymerases contain an alanine at this amino acid. In vitro polymerase assays demonstrated that HSV-1, HSV-2, HCMV, VZV, and HHV-8 polymerases were inhibited by PNU-183792, whereas the HHV-6 polymerase was not. Changing this amino acid from valine to alanine in the HSV-1, HCMV, and HHV-8 polymerases alters the polymerase activity so that it is less sensitive to drug inhibition. In contrast, changing the equivalent amino acid in the HHV-6 polymerase from alanine to valine alters polymerase activity so that PNU-183792 inhibits this enzyme. The HSV-1, HSV-2, and HCMV drug-resistant mutants were not altered in their susceptibilities to nucleoside analogs; in fact, some of the mutants were hypersensitive to several of the drugs. These results support a mechanism where PNU-183792 inhibits herpesviruses by interacting with a binding determinant on the viral DNA polymerase that is less important for the binding of nucleoside analogs and deoxynucleoside triphosphates.
Asunto(s)
Antivirales/farmacología , Citomegalovirus/efectos de los fármacos , ADN Polimerasa Dirigida por ADN/genética , Inhibidores Enzimáticos/farmacología , Herpesvirus Humano 1/efectos de los fármacos , Herpesvirus Humano 2/efectos de los fármacos , Mutación , Inhibidores de la Síntesis del Ácido Nucleico , Quinolinas/farmacología , Animales , Chlorocebus aethiops , Farmacorresistencia Viral , Humanos , Células VeroRESUMEN
Through broad screening of the compound library at Pharmacia, a naphthalene carboxamide was identified as a nonnucleoside inhibitor of human cytomegalovirus (HCMV) polymerase. Structure-activity relationship studies demonstrated that a quinoline ring could be substituted for naphthalene, resulting in the discovery of a 4-hydroxyquinoline-3-carboxamide (4-HQC) class of antiviral agents with unique biological properties. In vitro assays with the 4-HQCs have demonstrated potent inhibition of HCMV, herpes simplex virus type 1 (HSV-1), and varicella-zoster virus (VZV) polymerases but no inhibition of human alpha, delta, and gamma polymerases. Antiviral cell culture assays have further confirmed that these compounds are active against HCMV, HSV-1, HSV-2, VZV, and many animal herpesviruses. However, these compounds were not active against several nonherpesviruses representing different DNA and RNA virus families. A strong correlation between the viral DNA polymerase and antiviral activity for this class of compounds supports inhibition of the viral polymerase as the mechanism of antiviral activity. Northern blot analysis of immediate-early and late viral transcripts also pointed to a block in the viral life cycle consistent with inhibition of viral DNA replication. In vitro HCMV polymerase assays indicate that the 4-HQCs are competitive inhibitors of nucleoside binding. However, no cross-resistance could be detected with ganciclovir-resistant HCMV or acyclovir-resistant HSV-1 mutants. The unique, broad-spectrum activities of the 4-HQCs may offer new opportunities for treating many of the diseases caused by herpesviruses.