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Identification of A Novel Class of Benzofuran Oxoacetic Acid-Derived Ligands that Selectively Activate Cellular EPAC1
Cyclic AMP promotes EPAC1 and EPAC2 activation through direct binding to a specific cyclic nucleotide-binding domain (CNBD) within each protein, leading to activation of Rap GTPases, which control multiple cell responses, including cell proliferation, adhesion, morphology, exocytosis, and gene expre...
Autores principales: | , , , , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6912754/ https://www.ncbi.nlm.nih.gov/pubmed/31726720 http://dx.doi.org/10.3390/cells8111425 |
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author | M. Beck, Elizabeth Parnell, Euan Cowley, Angela Porter, Alison Gillespie, Jonathan Robinson, John Robinson, Lindsay Pannifer, Andrew D. Hamon, Veronique Jones, Philip Morrison, Angus McElroy, Stuart Timmerman, Martin Rutjes, Helma Mahajan, Pravin Wiejak, Jolanta Luchowska-Stańska, Urszula Morgan, David Barker, Graeme Rehmann, Holger Yarwood, Stephen J. |
author_facet | M. Beck, Elizabeth Parnell, Euan Cowley, Angela Porter, Alison Gillespie, Jonathan Robinson, John Robinson, Lindsay Pannifer, Andrew D. Hamon, Veronique Jones, Philip Morrison, Angus McElroy, Stuart Timmerman, Martin Rutjes, Helma Mahajan, Pravin Wiejak, Jolanta Luchowska-Stańska, Urszula Morgan, David Barker, Graeme Rehmann, Holger Yarwood, Stephen J. |
author_sort | M. Beck, Elizabeth |
collection | PubMed |
description | Cyclic AMP promotes EPAC1 and EPAC2 activation through direct binding to a specific cyclic nucleotide-binding domain (CNBD) within each protein, leading to activation of Rap GTPases, which control multiple cell responses, including cell proliferation, adhesion, morphology, exocytosis, and gene expression. As a result, it has become apparent that directed activation of EPAC1 and EPAC2 with synthetic agonists may also be useful for the future treatment of diabetes and cardiovascular diseases. To identify new EPAC agonists we have developed a fluorescent-based, ultra-high-throughput screening (uHTS) assay that measures the displacement of binding of the fluorescent cAMP analogue, 8-NBD-cAMP to the EPAC1 CNBD. Triage of the output of an approximately 350,000 compound screens using this assay identified a benzofuran oxaloacetic acid EPAC1 binder (SY000) that displayed moderate potency using orthogonal assays (competition binding and microscale thermophoresis). We next generated a limited library of 91 analogues of SY000 and identified SY009, with modifications to the benzofuran ring associated with a 10-fold increase in potency towards EPAC1 over SY000 in binding assays. In vitro EPAC1 activity assays confirmed the agonist potential of these molecules in comparison with the known EPAC1 non-cyclic nucleotide (NCN) partial agonist, I942. Rap1 GTPase activation assays further demonstrated that SY009 selectively activates EPAC1 over EPAC2 in cells. SY009 therefore represents a novel class of NCN EPAC1 activators that selectively activate EPAC1 in cellulae. |
format | Online Article Text |
id | pubmed-6912754 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-69127542020-01-02 Identification of A Novel Class of Benzofuran Oxoacetic Acid-Derived Ligands that Selectively Activate Cellular EPAC1 M. Beck, Elizabeth Parnell, Euan Cowley, Angela Porter, Alison Gillespie, Jonathan Robinson, John Robinson, Lindsay Pannifer, Andrew D. Hamon, Veronique Jones, Philip Morrison, Angus McElroy, Stuart Timmerman, Martin Rutjes, Helma Mahajan, Pravin Wiejak, Jolanta Luchowska-Stańska, Urszula Morgan, David Barker, Graeme Rehmann, Holger Yarwood, Stephen J. Cells Article Cyclic AMP promotes EPAC1 and EPAC2 activation through direct binding to a specific cyclic nucleotide-binding domain (CNBD) within each protein, leading to activation of Rap GTPases, which control multiple cell responses, including cell proliferation, adhesion, morphology, exocytosis, and gene expression. As a result, it has become apparent that directed activation of EPAC1 and EPAC2 with synthetic agonists may also be useful for the future treatment of diabetes and cardiovascular diseases. To identify new EPAC agonists we have developed a fluorescent-based, ultra-high-throughput screening (uHTS) assay that measures the displacement of binding of the fluorescent cAMP analogue, 8-NBD-cAMP to the EPAC1 CNBD. Triage of the output of an approximately 350,000 compound screens using this assay identified a benzofuran oxaloacetic acid EPAC1 binder (SY000) that displayed moderate potency using orthogonal assays (competition binding and microscale thermophoresis). We next generated a limited library of 91 analogues of SY000 and identified SY009, with modifications to the benzofuran ring associated with a 10-fold increase in potency towards EPAC1 over SY000 in binding assays. In vitro EPAC1 activity assays confirmed the agonist potential of these molecules in comparison with the known EPAC1 non-cyclic nucleotide (NCN) partial agonist, I942. Rap1 GTPase activation assays further demonstrated that SY009 selectively activates EPAC1 over EPAC2 in cells. SY009 therefore represents a novel class of NCN EPAC1 activators that selectively activate EPAC1 in cellulae. MDPI 2019-11-12 /pmc/articles/PMC6912754/ /pubmed/31726720 http://dx.doi.org/10.3390/cells8111425 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article M. Beck, Elizabeth Parnell, Euan Cowley, Angela Porter, Alison Gillespie, Jonathan Robinson, John Robinson, Lindsay Pannifer, Andrew D. Hamon, Veronique Jones, Philip Morrison, Angus McElroy, Stuart Timmerman, Martin Rutjes, Helma Mahajan, Pravin Wiejak, Jolanta Luchowska-Stańska, Urszula Morgan, David Barker, Graeme Rehmann, Holger Yarwood, Stephen J. Identification of A Novel Class of Benzofuran Oxoacetic Acid-Derived Ligands that Selectively Activate Cellular EPAC1 |
title | Identification of A Novel Class of Benzofuran Oxoacetic Acid-Derived Ligands that Selectively Activate Cellular EPAC1 |
title_full | Identification of A Novel Class of Benzofuran Oxoacetic Acid-Derived Ligands that Selectively Activate Cellular EPAC1 |
title_fullStr | Identification of A Novel Class of Benzofuran Oxoacetic Acid-Derived Ligands that Selectively Activate Cellular EPAC1 |
title_full_unstemmed | Identification of A Novel Class of Benzofuran Oxoacetic Acid-Derived Ligands that Selectively Activate Cellular EPAC1 |
title_short | Identification of A Novel Class of Benzofuran Oxoacetic Acid-Derived Ligands that Selectively Activate Cellular EPAC1 |
title_sort | identification of a novel class of benzofuran oxoacetic acid-derived ligands that selectively activate cellular epac1 |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6912754/ https://www.ncbi.nlm.nih.gov/pubmed/31726720 http://dx.doi.org/10.3390/cells8111425 |
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