Synthesis of novel (-)-epicatechin derivatives as potential endothelial GPER agonists: Evaluation of biological effects
Por:
Sarmiento V., Ramirez-Sanchez I., Moreno-Ulloa A., Romero-Perez D., Chávez D., Ortiz M., Najera N., Correa-Basurto J., Villarreal F., Ceballos G.
Publicada:
1 ene 2018
Resumen:
To potentially identify proteins that interact (i.e. bind) and may contribute to mediate (-)-epicatechin (Epi) responses in endothelial cells we implemented the following strategy: 1) synthesis of novel Epi derivatives amenable to affinity column use, 2) in silico molecular docking studies of the novel derivatives on G protein-coupled estrogen receptor (GPER), 3) biological assessment of the derivatives on NO production, 4) implementation of an immobilized Epi derivative affinity column and, 5) affinity column based isolation of Epi interacting proteins from endothelial cell protein extracts. For these purposes, the Epi phenol and C3 hydroxyl groups were chemically modified with propargyl or mesyl groups. Docking studies of the novel Epi derivatives on GPER conformers at 14 ns and 70 ns demostrated favorable thermodynamic interactions reaching the binding site. Cultures of bovine coronary artery endothelial cells (BCAEC) treated with Epi derivatives stimulated NO production via Ser1179 phosphorylation of eNOS, effects that were attenuated by the use of the GPER blocker, G15. Epi derivative affinity columns yielded multiple proteins from BCAEC. Proteins were electrophoretically separated and inmmunoblotting analysis revealed GPER as an Epi derivative binding protein. Altogether, these results validate the proposed strategy to potentially isolate and identify novel Epi receptors that may account for its biological activity. © 2018 Elsevier Ltd
Filiaciones:
Sarmiento V.:
Universidad Autónoma de Baja California, Tijuana, BC, Mexico
Ramirez-Sanchez I.:
Escuela Superior de Medicina del Instituto Politécnico Nacional, Sección de Posgrado, Mexico
Moreno-Ulloa A.:
Departamento de Innovación Biomédica, Centro de Investigación Científica y de, Educación Superior de Ensenada (CICESE)BC, Mexico
Romero-Perez D.:
Universidad Autónoma de Baja California, Tijuana, BC, Mexico
Chávez D.:
Centro de Graduados e Investigación en Química del Instituto Tecnológico de Tijuana, Apartado Postal 1166, Tijuana, BC 22510, Mexico
Ortiz M.:
Escuela Superior de Medicina del Instituto Politécnico Nacional, Sección de Posgrado, Mexico
Najera N.:
Escuela Superior de Medicina del Instituto Politécnico Nacional, Sección de Posgrado, Mexico
Correa-Basurto J.:
Escuela Superior de Medicina del Instituto Politécnico Nacional, Sección de Posgrado, Mexico
Villarreal F.:
University of California, San Diego, School of Medicine, United States
Ceballos G.:
Escuela Superior de Medicina del Instituto Politécnico Nacional, Sección de Posgrado, Mexico
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