RESUMEN
3-(1,1-Dioxo-2H-[1,2,4]benzothiadiazin-3-yl)-4-hydroxy-2H-quinolizin-2-one derivatives as potential anti-HCV drugs targeting NS5B polymerase have been investigated. Their synthesis, HCV NS5B polymerase inhibition, and replicon activity are discussed.
Asunto(s)
Antivirales/síntesis química , Benzotiadiazinas/síntesis química , Inhibidores Enzimáticos/síntesis química , Hepatitis C/tratamiento farmacológico , Quinolizinas/síntesis química , Proteínas no Estructurales Virales/antagonistas & inhibidores , Animales , Antivirales/farmacología , Benzotiadiazinas/farmacología , Química Farmacéutica/métodos , Diseño de Fármacos , Inhibidores Enzimáticos/farmacología , Hepacivirus/metabolismo , Técnicas In Vitro , Concentración 50 Inhibidora , Espectroscopía de Resonancia Magnética , Modelos Químicos , Estructura Molecular , Quinolizinas/farmacología , Relación Estructura-Actividad , Proteínas no Estructurales Virales/químicaRESUMEN
2-(1,1-Dioxo-2H-[1,2,4]benzothiadiazin-3-yl)-1-hydroxynaphthalene derivatives as potential anti-HCV drugs targeting NS5B polymerase have been investigated. Their synthesis, HCV NS5B polymerase inhibition and replicon activity are discussed.
Asunto(s)
Antivirales/síntesis química , Benzotiadiazinas/síntesis química , Inhibidores Enzimáticos/síntesis química , Hepatitis C/tratamiento farmacológico , Naftalenos/antagonistas & inhibidores , Naftoles/síntesis química , Proteínas no Estructurales Virales/antagonistas & inhibidores , Animales , Antivirales/farmacología , Benzotiadiazinas/farmacología , Química Farmacéutica/métodos , Diseño de Fármacos , Inhibidores Enzimáticos/farmacología , Hepacivirus/metabolismo , Humanos , Técnicas In Vitro , Concentración 50 Inhibidora , Espectroscopía de Resonancia Magnética , Modelos Químicos , Estructura Molecular , Naftalenos/química , Naftoles/farmacología , Relación Estructura-ActividadRESUMEN
A direct UV-VIS spectrophotometric assay has been developed for peptide deformylase. This assay employs a novel class of peptide mimetics as deformylase substrates which, upon enzymatic removal of the N-terminal formyl group, rapidly release free thiols. The released thiols are quantitated using Ellman's reagent. A variety of peptide analogues that contain beta-thiaphenylalanine or beta-thiamethionine as the N-terminal residue were synthesized and found to be excellent substrates of the peptide deformylase from Escherichia coli (k(cat)/K(M) = 6.9 x 10(5) M(-1) s(-1) for the most reactive substrate). The deformylase reaction is conveniently monitored on a UV-VIS spectrophotometer in a continuous fashion. The versatility of the assay has been demonstrated by its application to kinetic characterization of the deformylase, pH profile studies, and enzyme inhibition assays. The assay can also be performed in an end-point fashion. The results demonstrate that this assay is a simple, highly sensitive, and rapid method to study kinetic properties of deformylases without the use of any coupling enzymes.