RESUMEN
Selective bradykinin (BK) B 1 receptor antagonists could be novel therapeutic agents for the treatment of pain and inflammation. Elucidation of the structure activity relationships of the structurally novel HTS lead compound 1 provided potent hBK B 1 receptor antagonists with excellent receptor occupancy in the CNS of hBK B 1 transgenic rats.
Asunto(s)
Aminas/química , Benzofenonas/química , Benzofenonas/farmacología , Antagonistas del Receptor de Bradiquinina B1 , Animales , Benzofenonas/síntesis química , Línea Celular , Perros , Humanos , Estructura Molecular , Ratas , Receptor de Bradiquinina B1/metabolismo , Relación Estructura-ActividadRESUMEN
Selective bradykinin (BK) B(1) receptor antagonists have been shown to be antinociceptive in animal models and could be novel therapeutic agents for the treatment of pain and inflammation. Elucidation of the structure-activity relationships of the biphenyl moiety of the lead compound 1 provided a potent new structural class of BK B(1) receptor antagonists.
Asunto(s)
Analgésicos/química , Antiinflamatorios no Esteroideos/química , Antagonistas del Receptor de Bradiquinina B1 , Ciclohexanos/química , Hidrocarburos Fluorados/química , Piridinas/química , Analgésicos/síntesis química , Analgésicos/farmacología , Animales , Animales Modificados Genéticamente , Antiinflamatorios no Esteroideos/síntesis química , Antiinflamatorios no Esteroideos/farmacología , Ciclohexanos/síntesis química , Ciclohexanos/farmacología , Humanos , Hidrocarburos Fluorados/síntesis química , Hidrocarburos Fluorados/farmacología , Piridinas/síntesis química , Piridinas/farmacología , Ratas , Receptor de Bradiquinina B1/genética , Relación Estructura-ActividadRESUMEN
We report the critical residues for the interaction of the kinins with human bradykinin receptor 1 (B1) using site-directed mutagenesis in conjunction with molecular modeling of the binding modes of the kinins in the homology model of the B1 receptor. Mutation of Lys118 in transmembrane (TM) helix 3, Ala270 in TM6, and Leu294 in TM7 causes a significant decrease in the affinity for the peptide agonists des-Arg10kallidin (KD) and des-Arg9BK but not the peptide antagonist des-Arg10Leu9KD. In contrast, mutations in TM2, TM3, TM6, and TM7 cause a significant decrease in the affinity for both the peptide agonists and the antagonist. These data indicate that the B1 bradykinin binding pocket for agonists and antagonists is similar, but the manners in which they interact with the receptor do not completely overlap. Therefore, there is a potential to influence the receptor's ligand selectivity.
Asunto(s)
Cininas/química , Cininas/metabolismo , Modelos Moleculares , Receptor de Bradiquinina B1/química , Receptor de Bradiquinina B1/metabolismo , Secuencia de Aminoácidos , Animales , Sitios de Unión , Membrana Celular/química , Membrana Celular/metabolismo , Secuencia Conservada , Humanos , Datos de Secuencia Molecular , Mutagénesis Sitio-Dirigida , Mutación/genética , Unión Proteica , Estructura Cuaternaria de Proteína , Estructura Secundaria de Proteína , Receptor de Bradiquinina B1/genética , Alineación de SecuenciaRESUMEN
We report the first homology model of human bradykinin receptor B1 generated from the crystal structure of bovine rhodopsin as a template. Using an automated docking procedure, two B1 receptor antagonists of the dihydroquinoxalinone structural class were docked into the receptor model. Site-directed mutagenesis data of the amino acid residues in TM1, TM3, TM6, and TM7 were incorporated to place the compounds in the binding site of the homology model of the human B1 bradykinin receptor. The best pose in agreement with the mutation data was selected for detailed study of the receptor-antagonist interaction. To test the model, the calculated antagonist-receptor binding energy was correlated with the experimentally measured binding affinity (K(i)) for nine dihydroquinoxalinone analogs. The model was used to gain insight into the molecular mechanism for receptor function and to optimize the dihydroquinoxalinone analogs.
Asunto(s)
Modelos Moleculares , Quinoxalinas/química , Receptor de Bradiquinina B1/química , Secuencia de Aminoácidos , Sitios de Unión , Humanos , Datos de Secuencia Molecular , Mutagénesis Sitio-Dirigida , Unión Proteica , Estructura Secundaria de Proteína , Receptor de Bradiquinina B1/genética , Receptor de Bradiquinina B1/metabolismo , Rodopsina/química , Alineación de Secuencia , Homología Estructural de ProteínaRESUMEN
We have developed an efficient and selective radioligand, the [35S]-radiolabeled dihydroquinoxalinone derivative, 4, for an ex vivo receptor occupancy assay in transgenic rats over-expressing the human bradykinin B1 receptor.