RESUMEN
Malonic acid hydroxamate derivatives bis-substituted at the methylene group were synthesized as potential nonpeptidic inhibitors of human neutrophil collagenase (MMP8). The presence of an aromatic residue both at the C2 malonic acid position and in the C-terminal tail for hydrophobic interactions with the surface-exposed S1 binding site and the S1' pocket of the enzyme, respectively, was found to be sufficient for submicromolar inhibition potencies. For optimal insertion of the aryl amide group into the hydrophobic S1' pocket, spacing of the C-terminal phenyl group by at least a 3C-chain was required. In view of these results the achiral indan-2, 2-dicarboxylic acid was used to mimic the 2-benzyl-2-methylmalonic acid residue, and its derivatization to the 3-phenylpropyl amide hydroxamate produced a potent, achiral, low-mass inhibitor of MMP8 (Ki = 0.3 microM), the binding mode of which was unambiguously determined by X-ray crystallographic analysis.
Asunto(s)
Inhibidores Enzimáticos , Ácidos Hidroxámicos , Malonatos , Inhibidores de la Metaloproteinasa de la Matriz , Sitios de Unión , Colagenasas/metabolismo , Cristalografía por Rayos X , Inhibidores Enzimáticos/síntesis química , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/metabolismo , Inhibidores Enzimáticos/farmacología , Humanos , Ácidos Hidroxámicos/síntesis química , Ácidos Hidroxámicos/química , Ácidos Hidroxámicos/metabolismo , Ácidos Hidroxámicos/farmacología , Malonatos/síntesis química , Malonatos/química , Malonatos/metabolismo , Malonatos/farmacología , Metaloproteinasa 8 de la Matriz , Modelos Moleculares , Estereoisomerismo , Relación Estructura-ActividadRESUMEN
For most of the known synthetic inhibitors of matrix metalloproteinases (MMPs), a substrate-like binding mode was postulated on the basis of X-ray crystallographic structures of MMP/inhibitor complexes. Conversely, the malonic acid-based inhibitor (2R,S)-HONH-CO-CH(i-Bu)-CO-Ala-Gly-NH2 was found to bind in a surprisingly different manner. Using this compound as a new lead structure, the interaction sites with human neutrophil collagenase (MMP8) were optimized with a series of iteratively designed analogues and with the help of X-ray structural analysis of selected inhibitors to finally produce low molecular weight nonpeptidic compounds of 500-1000-fold improved inhibitory potency.
Asunto(s)
Diseño de Fármacos , Inhibidores Enzimáticos/síntesis química , Malonatos/química , Inhibidores de la Metaloproteinasa de la Matriz , Cristalografía por Rayos X , Humanos , Espectroscopía de Resonancia Magnética , Metaloproteinasa 8 de la Matriz , Estructura Molecular , Espectrometría de Masa Bombardeada por Átomos VelocesRESUMEN
A method has been developed for the assignment of HPLC peaks to their corresponding compounds in libraries of single compounds (parallel syntheses). The basis of the new method is the correlation of the product retention times with the different substituents in the variable positions of the molecule. The correlation is performed automatically by a new algorithm which is part of the computer program LIBFINDER. This practical, easy-to-use tool accelerates the analysis, characterization and purification of chemical libraries, without the need for expensive HPLC-MS equipment.