RESUMEN
A novel series of imidazolidinone-based matrix metalloproteinase (MMP) inhibitors was discovered by structural modification of pyrrolidinone la. Potent inhibition of MMP-13 was exhibited by the analogues having 4-(4-fluorophenoxy)phenyl (4a, IC50 = 3 nM) and 4-(naphth-2-yloxy)phenyl (4h, IC50 = 4 nM) as P1' groups.
Asunto(s)
Amidas/síntesis química , Amidas/farmacología , Ácidos Carboxílicos/síntesis química , Ácidos Carboxílicos/farmacología , Inhibidores Enzimáticos/síntesis química , Imidazoles/síntesis química , Imidazoles/farmacología , Inhibidores de la Metaloproteinasa de la Matriz , Diseño de Fármacos , Inhibidores Enzimáticos/farmacología , Metaloproteinasa 13 de la Matriz , EstereoisomerismoAsunto(s)
Bencenoacetamidas , Ácidos Hidroxámicos/síntesis química , Inhibidores de la Metaloproteinasa de la Matriz , Inhibidores de Proteasas/síntesis química , Pirrolidinonas/síntesis química , Colagenasas/química , Diseño de Fármacos , Ácidos Hidroxámicos/química , Metaloproteinasa 13 de la Matriz , Modelos Moleculares , Inhibidores de Proteasas/química , Pirrolidinonas/química , Relación Estructura-ActividadAsunto(s)
Cartílago Articular/enzimología , Colagenasas/genética , Regulación de la Expresión Génica , Animales , Cartílago Articular/citología , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Interleucina-1/farmacología , Articulaciones , Metaloproteinasa 1 de la Matriz , Metaloproteinasa 13 de la Matriz , ARN Mensajero/genética , Conejos , Proteínas Recombinantes/farmacología , Transcripción Genética/efectos de los fármacosRESUMEN
Through the use of empirical and computational methods, phosphinate-based inhibitors of MMP-1 and MMP-13 that bind into the S2 pocket of these enzymes were designed. The synthesis and testing of 2 suggested that binding was occurring as hypothesized. Structure determination of a co-crystal of 2 bound to the catalytic domain of MMP-1 confirmed the binding mode. Substituents binding into S2, S1', S2' and S3', were optimized yielding compounds with low double-digit nM IC50's against these enzymes.
Asunto(s)
Inhibidores de la Metaloproteinasa de la Matriz , Ácidos Fosfínicos/farmacología , Sitios de Unión , Colagenasas/farmacocinética , Simulación por Computador , Cristalografía por Rayos X , Diseño de Fármacos , Concentración 50 Inhibidora , Metaloproteinasa 1 de la Matriz , Metaloproteinasa 13 de la Matriz , Modelos MolecularesRESUMEN
Proteolysis of triple-helical collagen is an important step in the progression toward irreversible tissue damage in osteoarthritis. Earlier work on the expression of enzymes in cartilage suggested that collagenase-1 (MMP-1) contributes to the process. Degenerate reverse transcription polymerase chain reaction experiments, Northern blot analysis, and direct immunodetection have now provided evidence that collagenase-3 (MMP-13), an enzyme recently cloned from human breast carcinoma, is expressed by chondrocytes in human osteoarthritic cartilage. Variable levels of MMP-13 and MMP-1 in cartilage was significantly induced at both the message and protein levels by interleukin-1 alpha. Recombinant MMP-13 cleaved type II collagen to give characteristic 3/4 and 1/4 fragments; however, MMP-13 turned over type II collagen at least 10 times faster than MMP-1. Experiments with intact type II collagen as well as a synthetic peptide suggested that MMP-13 cleaved type II collagen at the same bond as MMP-1, but this was then followed by a secondary cleavage that removed three amino acids from the 1/4 fragment amino terminus. The expression of MMP-13 in osteoarthritic cartilage and its activity against type II collagen suggest that the enzyme plays a significant role in cartilage collagen degradation, and must consequently form part of a complex target for proposed therapeutic interventions based on collagenase inhibition.
Asunto(s)
Cartílago/enzimología , Colágeno/metabolismo , Colagenasas/metabolismo , Osteoartritis/enzimología , Secuencia de Aminoácidos , Secuencia de Bases , Clonación Molecular , Colagenasas/genética , Humanos , Cinética , Metaloproteinasa 13 de la Matriz , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , Análisis de Secuencia , Homología de Secuencia de Aminoácido , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Especificidad por SustratoRESUMEN
Interleukin-1 alpha (IL-1) stimulated the release of degraded proteoglycan from primary cultures of chondrocyte monolayers in a time- and dose-dependent fashion. Bafilomycin A1, a specific inhibitor of the vacuolar H(+)-ATPase, efficiently blocked acidification of the chondrocyte vacuolar system. Under these conditions IL-1-stimulated proteoglycan degradation was inhibited by bafilomycin A1 with an IC50 of < 10 nM in both chondrocyte monolayers and articular cartilage explants. This concentration was at least 100-fold less than that required to partially inhibit total protein synthesis. In chondrocyte monolayers, bafilomycin A1 could be added several hours after IL-1 and complete inhibition was still observed. Tumor necrosis factor-alpha and retinoic acid also stimulated proteoglycan degradation in chondrocyte monolayers, and in both cases the response was inhibited by bafilomycin A1. These results suggest that maintenance of vacuolar acidity is required for cytokine stimulated proteoglycan degradation and that this requirement is at a point distal to receptor binding and early signal transduction events. IL-1 also stimulated the synthesis and secretion of prostromelysin by chondrocyte monolayers, however, under conditions in which IL-1 stimulated proteoglycan release was totally blocked by bafilomycin A1, there was no effect on IL-1-stimulated stromelysin secretion or stromelysin enzyme activity. These results, in which stromelysin synthesis and proteoglycan degradation were dissociated, suggest that an additional enzyme is responsible for proteoglycan degradation in this chondrocyte monolayer system.