RESUMEN
G-protein-coupled receptors (GPCRs) play a major role in psychiatric disorders and are the targets of several current therapeutic approaches in this field. A number of studies have now shown that GPCRs can assemble as high molecular weight homo- and hetero-oligomers, which could affect ligand binding, intracellular signalling or trafficking. This information could be critical in design of new drugs to treat neurological and psychiatric disorders. This chapter describes a sequential co-immunoprecipitation and immunoblot protocol for determining oligomerisation of the 5-hydroxytryptamine (HT)1A receptor with other GPCRs in co-transfected HEK-293 cells.
Asunto(s)
Western Blotting/métodos , Cromatografía de Afinidad/métodos , Inmunoprecipitación/métodos , Receptores Acoplados a Proteínas G/análisis , Células HEK293 , Humanos , Oligopéptidos , Multimerización de Proteína , Proteínas Proto-Oncogénicas c-myc , Receptor de Serotonina 5-HT1A/análisis , Receptor de Serotonina 5-HT1A/química , Receptor de Serotonina 5-HT1B/análisis , Receptor de Serotonina 5-HT1B/química , Receptor de Serotonina 5-HT1D/análisis , Receptor de Serotonina 5-HT1D/química , Receptores Acoplados a Proteínas G/química , Proteínas Recombinantes de Fusión/análisis , Proteínas Recombinantes de Fusión/química , TransfecciónRESUMEN
A series of 3-[3-(4-aryl-1-piperazinyl)-propyl]-1H-indole derivatives (12a-h) was synthesized and evaluated for binding affinity at the human 5-hydroxytryptamine(1A) receptor (5-HT(1A)R) compounds (12b) and (12h) showed the highest 5-HT(1A) receptor affinity (IC(50)=15 nM). Molecular docking studies with all the compounds in a homology model of 5-HT(1A) showed that the main interaction anchoring the ligand in the receptor was a charge-reinforced bond between the protonated nitrogen atom (N-4) of the piperazine ring and Aspartate(3.32).
Asunto(s)
Indoles/química , Piperazinas/química , Receptor de Serotonina 5-HT1A/química , Ácido Aspártico/química , Sitios de Unión , Simulación por Computador , Humanos , Indoles/síntesis química , Piperazina , Estructura Terciaria de Proteína , Receptor de Serotonina 5-HT1A/metabolismo , Relación Estructura-ActividadRESUMEN
Arylpiperazine compounds are promising 5-HT(1A) receptor ligands that can contribute for accelerating the onset of therapeutic effect of selective serotonin reuptake inhibitors. In the present work, the chemometric methods HCA, PCA, KNN, SIMCA and PLS were employed in order to obtain SAR and QSAR models relating the structures of arylpiperazine compounds to their 5-HT(1A) receptor affinities. A training set of 52 compounds was used to construct the models and the best ones were obtained with nine topological descriptors. The classification and regression models were externally validated by means of predictions for a test set of 14 compounds and have presented good quality, as verified by the correctness of classifications, in the case of pattern recognition studies, and by the high correlation coefficients (q(2)=0.76, r(2)=0.83) and small prediction errors for the PLS regression. Since the results are in good agreement with previous SAR studies, we can suggest that these findings can help in the search for 5-HT(1A) receptor ligands that are able to improve antidepressant treatment.