Diseño racional de inhibidores de la actividad RGS de GRK2

GRK2 overexpression is involved in the development of several pathologies with unsatisfied therapeutic demand. Plenty evidences demonstrate a desensitizating mechanism for GPCRs, associated to GRK2 RGS homology domain, and independent of its kinase domain. Since GRK2 potential as therapeutic target...

Descripción completa

Guardado en:
Detalles Bibliográficos
Autor principal: Echeverría, Emiliana
Otros Autores: Fernández, Natalia
Formato: Tesis doctoral acceptedVersion
Lenguaje:Español
Publicado: Facultad de Farmacia y Bioquímica 2018
Materias:
GRK2
Docking
Desensibilización
GPCR
Ciencias de la vida
Acceso en línea:http://repositoriouba.sisbi.uba.ar/gsdl/cgi-bin/library.cgi?a=d&c=posgraafa&cl=CL1&d=HWA_5773
http://repositoriouba.sisbi.uba.ar/gsdl/collect/posgraafa/index/assoc/HWA_5773.dir/5773.PDF
Aporte de:
Repositorio Digital de la Universidad de Buenos Aires (UBA) de Universidad de Buenos Aires
id I28-R145-HWA_5773
record_format dspace
spelling I28-R145-HWA_57732022-03-03 GRK2 overexpression is involved in the development of several pathologies with unsatisfied therapeutic demand. Plenty evidences demonstrate a desensitizating mechanism for GPCRs, associated to GRK2 RGS homology domain, and independent of its kinase domain. Since GRK2 potential as therapeutic target could lie on its RGS activity, we performed a virtual screening of Enamine library, setting a docking on its RH domain. From 13 compounds (C1-C13) with acceptable in silico binding energies, 4 exerted in vitro activity. C2, C3, C4 and C5 potentiated H2r cAMP response, inhibited GRK2 membrane translocation induced by receptor overexpression, and inhibited coinmunoprecipitation of GRK2 with G?s. We evaluated preliminarily their ADMET properties, and observed that all 4 hits have a suitable lipophilicity, but C3 and C5 stood out, as they didn´t affect cellular viability. After discarding C5 because it also potentiated H1r calcium response, C3 was defined as prototype for analogs design. From those analogs synthesized and evaluated in vitro, we found one that inhibited RGS GRK2 activity by potentiating H2r AMPc response. This compounds could be useful in elucidating other RH functions, and in the future, they could lay the foundation for pharmacologic therapy of those diseases where GRK2 activity is exacerbated. Fil: Echeverría, Emiliana. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Buenos Aires, Argentina Facultad de Farmacia y Bioquímica Fernández, Natalia Echeverría, Emiliana 2018-03-28 La sobreexpresión de GRK2 está asociada al desarrollo de numerosas patologías con una demanda terapéutica insatisfecha. Evidencias demuestran que su dominio homólogo a RGS media la desensibilización de GPCRs independientemente de su fosforilación. Como el potencial de GRK2 como blanco terapéutico podría residir en su actividad RGS, realizamos un screeningvirtualde la biblioteca de Enamine, centrando undocking en su dominio RH. De trece compuestos (C1-C13) con buenas energías de unión in silico, cuatro presentaron actividad in vitro. C2, C3, C4 y C5 potenciaron la respuesta de AMPc del rH2, inhibieron la translocación a membrana de GRK2 inducida por la sobreexpresión del receptor, e inhibieron la coinmunoprecipitación entre GRK2 y G?s. Como evaluación preliminar de propiedades ADMET, observamos que todos los compuestos presentaron una lipofilia adecuada, pero C3 y C5 se destacaron por no afectar la viabilidad celular. Tras descartar C5, por potenciar también la respuesta de calcio del rH1, C3 fue empleado como prototipo para diseñar análogos. De los seleccionados para su síntesis y evaluación biológica, uno inhibió la actividad RGS de GRK2 al potenciar la respuesta de AMPc del rH2. Estos inhibidores serán de utilidad para estudiar las funciones del RH de GRK2, y a futuro podrían sentar las bases para la terapia farmacológica de patologías debidas a una actividad exacerbada de GRK2. application/pdf Cremaschi, Graciela Coso, Omar Talevi, Alan GRK2 Docking Desensibilización GPCR spa Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by-nc-nd/2.5/ar/ Ciencias de la vida Diseño racional de inhibidores de la actividad RGS de GRK2 info:eu-repo/semantics/doctoralThesis info:ar-repo/semantics/tesis doctoral info:eu-repo/semantics/acceptedVersion http://repositoriouba.sisbi.uba.ar/gsdl/cgi-bin/library.cgi?a=d&c=posgraafa&cl=CL1&d=HWA_5773 http://repositoriouba.sisbi.uba.ar/gsdl/collect/posgraafa/index/assoc/HWA_5773.dir/5773.PDF
institution Universidad de Buenos Aires
institution_str I-28
repository_str R-145
collection Repositorio Digital de la Universidad de Buenos Aires (UBA)
language Español
orig_language_str_mv spa
topic GRK2
Docking
Desensibilización
GPCR
Ciencias de la vida
spellingShingle GRK2
Docking
Desensibilización
GPCR
Ciencias de la vida
Echeverría, Emiliana
Diseño racional de inhibidores de la actividad RGS de GRK2
topic_facet GRK2
Docking
Desensibilización
GPCR
Ciencias de la vida
description GRK2 overexpression is involved in the development of several pathologies with unsatisfied therapeutic demand. Plenty evidences demonstrate a desensitizating mechanism for GPCRs, associated to GRK2 RGS homology domain, and independent of its kinase domain. Since GRK2 potential as therapeutic target could lie on its RGS activity, we performed a virtual screening of Enamine library, setting a docking on its RH domain. From 13 compounds (C1-C13) with acceptable in silico binding energies, 4 exerted in vitro activity. C2, C3, C4 and C5 potentiated H2r cAMP response, inhibited GRK2 membrane translocation induced by receptor overexpression, and inhibited coinmunoprecipitation of GRK2 with G?s. We evaluated preliminarily their ADMET properties, and observed that all 4 hits have a suitable lipophilicity, but C3 and C5 stood out, as they didn´t affect cellular viability. After discarding C5 because it also potentiated H1r calcium response, C3 was defined as prototype for analogs design. From those analogs synthesized and evaluated in vitro, we found one that inhibited RGS GRK2 activity by potentiating H2r AMPc response. This compounds could be useful in elucidating other RH functions, and in the future, they could lay the foundation for pharmacologic therapy of those diseases where GRK2 activity is exacerbated.
author2 Fernández, Natalia
author_facet Fernández, Natalia
Echeverría, Emiliana
format Tesis doctoral
Tesis doctoral
acceptedVersion
author Echeverría, Emiliana
author_sort Echeverría, Emiliana
title Diseño racional de inhibidores de la actividad RGS de GRK2
title_short Diseño racional de inhibidores de la actividad RGS de GRK2
title_full Diseño racional de inhibidores de la actividad RGS de GRK2
title_fullStr Diseño racional de inhibidores de la actividad RGS de GRK2
title_full_unstemmed Diseño racional de inhibidores de la actividad RGS de GRK2
title_sort diseño racional de inhibidores de la actividad rgs de grk2
publisher Facultad de Farmacia y Bioquímica
publishDate 2018
url http://repositoriouba.sisbi.uba.ar/gsdl/cgi-bin/library.cgi?a=d&c=posgraafa&cl=CL1&d=HWA_5773
http://repositoriouba.sisbi.uba.ar/gsdl/collect/posgraafa/index/assoc/HWA_5773.dir/5773.PDF
work_keys_str_mv AT echeverriaemiliana disenoracionaldeinhibidoresdelaactividadrgsdegrk2
_version_ 1766017548683313152

Ejemplares similares