RESUMO
Based on X-ray crystallographic analysis of a peroxisome proliferator-activated receptor (PPAR) α/δ dual agonist complexed with human PPARs ligand binding domain (LBD), we previously reported the design and synthesis of a pyrene-based fluorescent PPARα/δ co-agonist 2. Here, we found that the fluorescence intensity of 2 increased upon binding to hPPARα-LBD, in a manner dependent upon the concentration of the LBD. But, surprisingly, the fluorescence intensity of 2 decreased concentration-dependently upon binding to hPPRδ-LBD. Site-directed mutagenesis of the two hPPAR subtypes clearly indicated that Trp264 of hPPARδ-LBD, located between H2' helix and H3 helix (omega loop), is critical for the concentration-dependent decrease in fluorescence intensity, which is suggested to be due to fluorescence resonance energy transfer (FRET) from the pyrene moiety of bound 2 to the nearby side-chain indole moiety of Trp264 in the hPPARδ-LBD.
Assuntos
PPAR alfa/agonistas , PPAR delta/agonistas , Fenilpropionatos/química , Fenilpropionatos/metabolismo , Pirenos/química , Pirenos/metabolismo , Sequência de Aminoácidos , Sítios de Ligação , Desenho de Fármacos , Fluorescência , Transferência Ressonante de Energia de Fluorescência , Humanos , Ligantes , Modelos Moleculares , Dados de Sequência Molecular , Mutagênese Sítio-Dirigida , PPAR alfa/genética , PPAR alfa/metabolismo , PPAR delta/genética , PPAR delta/metabolismo , Fenilpropionatos/farmacologia , Conformação Proteica , Pirenos/farmacologia , TriptofanoRESUMO
Peroxisome proliferator-activated receptor gamma (PPARγ) is a ligand-mediated transcription factor with roles in glucose, lipid, and lipoprotein homeostasis, and PPARγ ligands are expected have therapeutic potential in these as well as other areas. We report here the design, synthesis, crystallographic analysis, and computational studies of α-benzylphenylpropanoic acid PPARγ agonists. Interestingly, these compounds show a reversal of the stereochemistry-transactivation activity relationship observed with other phenylpropanoic acid ligands.
Assuntos
PPAR gama/agonistas , Fenilpropionatos/síntese química , Sítios de Ligação , Desenho de Fármacos , Células HEK293 , Humanos , Modelos Moleculares , Conformação Molecular , PPAR gama/química , PPAR gama/genética , Fenilpropionatos/química , Fenilpropionatos/farmacologia , Estereoisomerismo , Relação Estrutura-Atividade , Ativação TranscricionalRESUMO
Screening of our library of peroxisome proliferator-activated receptor (PPAR) agonists yielded several phenylpropanoic acid-derived gamma-secretase inhibitors (GSIs). Structure-activity relationship studies indicated that (R)-configuration of alpha-substituted phenylpropanoic acid structure and cinnamic acid structure is favorable to prepare Notch-sparing GSIs.