RESUMEN
The optimisation of a new series of high potency muscarinic M3 antagonists, derived from high throughput screening library hit is described.
Asunto(s)
Receptor Muscarínico M3/antagonistas & inhibidores , Compuestos de Espiro/química , Animales , Evaluación Preclínica de Medicamentos , Hepatocitos/metabolismo , Humanos , Microsomas/metabolismo , Ratas , Receptor Muscarínico M3/metabolismo , Compuestos de Espiro/síntesis química , Compuestos de Espiro/farmacologíaRESUMEN
The ATP-gated P2X(7) receptor (P2X(7)R) is a promising therapeutic target in chronic inflammatory diseases with highly specific antagonists currently under clinical trials for rheumatoid arthritis. Anti-inflammatory actions of P2X(7)R antagonists are considered to result from inhibition of P2X(7)R-induced release of proinflammatory cytokines from activated macrophages. However, P2X(7)Rs are also expressed in resting macrophages, suggesting that P2X(7)R may also signal via cytokine-independent mechanisms involved in joint disease. In this study, we examined P2X(7)R function in resting human lung macrophages and mouse bone marrow-derived macrophages and found that ATP induced rapid release of the lysosomal cysteine proteases cathepsin B, K, L, and S and that was independent of the presence of the proinflammatory cytokines IL-1beta and IL-18. Cathepsins released into the medium were effective to degrade collagen extracellular matrix. ATP-induced cathepsin release was abolished by P2X(7)R antagonists, absent from P2X(7)R(-/-) mouse macrophages, and not associated with cell death. Our results suggest P2X(7)R activation may play a novel and direct role in tissue damage through release of cathepsins independently of its proinflammatory actions via IL-1 cytokines.
Asunto(s)
Catepsinas/metabolismo , Citocinas/metabolismo , Artropatías/metabolismo , Macrófagos/metabolismo , Receptores Purinérgicos P2/metabolismo , Adenosina Trifosfato/farmacología , Animales , Western Blotting , Catepsina B/metabolismo , Células Cultivadas , Relación Dosis-Respuesta a Droga , Ensayo de Inmunoadsorción Enzimática , Etanercept , Humanos , Inmunoglobulina G/farmacología , Inmunosupresores/farmacología , Interleucina-1beta/metabolismo , Artropatías/genética , Células L , Macrófagos/citología , Macrófagos/efectos de los fármacos , Metotrexato/farmacología , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Estructura Molecular , Antagonistas del Receptor Purinérgico P2 , Receptores Purinérgicos P2/genética , Receptores Purinérgicos P2X7 , Receptores del Factor de Necrosis Tumoral , Tiazoles/química , Tiazoles/farmacologíaRESUMEN
A novel series of antagonists of the human P2X7 receptor is described. Modification of substituents enabled identification of compounds selective for the rat P2X7 receptor and provides useful pharmacological tools for evaluation of the role of P2X7 in disease.
