RESUMEN

A series of dipeptide nitriles with a thienyl alanine in P2 were identified as potent and selective cathepsin C inhibitors. Incorporation of a substituted cyclopropyl moiety in P1 effectively protects these derivatives against hydrolase activity in whole blood.

Asunto(s)

Catepsina C/antagonistas & inhibidores , Catepsina C/metabolismo , Dipéptidos/química , Dipéptidos/farmacología , Nitrilos/química , Nitrilos/farmacología , Animales , Línea Celular , Dipéptidos/sangre , Dipéptidos/síntesis química , Humanos , Nitrilos/sangre , Nitrilos/síntesis química , Ratas , Relación Estructura-Actividad

RESUMEN

The measurement of prostaglandin E synthase (PGES) activity is cumbersome because the product of the reaction, PGE(2), is not readily quantitated by spectral means. The activity of isolated PGES is typically determined by PGE(2) immunoassay or by high-performance liquid chromatography using radiolabeled substrate. A relatively rapid continuous spectrophotometric assay which uses 15-hydroxyprostaglandin dehydrogenase (PGDH) to couple the oxidation of the 15-hydroxy group of PGE(2) to the formation of NADH was developed. PGDH is relatively specific for PGE(2) over the substrate for the PGES reaction, PGH(2), allowing a highly reproducible assay of PGES activity to be obtained.

Asunto(s)

Oxidorreductasas Intramoleculares/metabolismo , Espectrofotometría/métodos , Dinoprostona/metabolismo , Hidroxiprostaglandina Deshidrogenasas/metabolismo , Oxidorreductasas Intramoleculares/análisis , Cinética , NAD/análisis , NAD/biosíntesis , Prostaglandina H2 , Prostaglandina-E Sintasas , Prostaglandinas H/metabolismo , Proteínas Recombinantes de Fusión/metabolismo , Reproducibilidad de los Resultados , Sensibilidad y Especificidad