A yeast-based chemical screen identifies a PDE inhibitor that elevates steroidogenesis in mouse Leydig cells via PDE8 and PDE4 inhibition
A cell-based high-throughput screen (HTS) was developed to detect phosphodiesterase 8 (PDE8) and PDE4/8 combination inhibitors. By replacing the Schizosaccharomyces pombe PDE gene with the murine PDE8A1 gene in strains lacking adenylyl cyclase, we generated strains whose protein kinase A (PKA)-stimu...
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| creator | Demirbas, Didem Wyman, Arlene R Shimizu-Albergine, Masami Cakici, Ozgur Beavo, Joseph A Hoffman, Charles S |
| description | A cell-based high-throughput screen (HTS) was developed to detect phosphodiesterase 8 (PDE8) and PDE4/8 combination inhibitors. By replacing the Schizosaccharomyces pombe PDE gene with the murine PDE8A1 gene in strains lacking adenylyl cyclase, we generated strains whose protein kinase A (PKA)-stimulated growth in 5-fluoro orotic acid (5FOA) medium reflects PDE8 activity. From our previously-identified PDE4 and PDE7 inhibitors, we identified a PDE4/8 inhibitor that allowed us to optimize screening conditions. Of 222,711 compounds screened, ∼0.2% displayed composite Z scores of >20. Additional yeast-based assays using the most effective 367 compounds identified 30 candidates for further characterization. Among these, compound BC8-15 displayed the lowest IC₅₀ value for both PDE4 and PDE8 inhibition in in vitro enzyme assays. This compound also displays significant activity against PDE10A and PDE11A. BC8-15 elevates steroidogenesis in mouse Leydig cells as a single pharmacological agent. Assays using BC8-15 and two structural derivatives support a model in which PDE8 is a primary regulator of testosterone production by Leydig cells, with an additional role for PDE4 in this process. BC8-15, BC8-15A, and BC8-15C, which are commercially available compounds, display distinct patterns of activity against PDE4, PDE8, PDE10A, and PDE11A, representing a chemical toolkit that could be used to examine the biological roles of these enzymes in cell culture systems. |
| doi_str_mv | 10.1371/journal.pone.0071279 |
| format | Article |
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By replacing the Schizosaccharomyces pombe PDE gene with the murine PDE8A1 gene in strains lacking adenylyl cyclase, we generated strains whose protein kinase A (PKA)-stimulated growth in 5-fluoro orotic acid (5FOA) medium reflects PDE8 activity. From our previously-identified PDE4 and PDE7 inhibitors, we identified a PDE4/8 inhibitor that allowed us to optimize screening conditions. Of 222,711 compounds screened, ∼0.2% displayed composite Z scores of >20. Additional yeast-based assays using the most effective 367 compounds identified 30 candidates for further characterization. Among these, compound BC8-15 displayed the lowest IC₅₀ value for both PDE4 and PDE8 inhibition in in vitro enzyme assays. This compound also displays significant activity against PDE10A and PDE11A. BC8-15 elevates steroidogenesis in mouse Leydig cells as a single pharmacological agent. Assays using BC8-15 and two structural derivatives support a model in which PDE8 is a primary regulator of testosterone production by Leydig cells, with an additional role for PDE4 in this process. BC8-15, BC8-15A, and BC8-15C, which are commercially available compounds, display distinct patterns of activity against PDE4, PDE8, PDE10A, and PDE11A, representing a chemical toolkit that could be used to examine the biological roles of these enzymes in cell culture systems.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0071279</identifier><identifier>PMID: 23967182</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Adenosine ; Animals ; Assaying ; Biological activity ; Biology ; Cell culture ; Cell Line, Tumor ; Cloning ; Cyclic Nucleotide Phosphodiesterases, Type 4 - chemistry ; Cyclic Nucleotide Phosphodiesterases, Type 4 - metabolism ; Drug Evaluation, Preclinical - methods ; Enzymes ; Gene expression ; Genetics ; Glucose ; High-Throughput Screening Assays ; Humans ; Inhibition ; Inhibitors ; Laboratory animals ; Leydig cells ; Leydig Cells - drug effects ; Leydig Cells - metabolism ; Lymphocytes ; Male ; Medical screening ; Mice ; Molecular Docking Simulation ; Orotic acid ; Pharmacology ; Phosphodiesterase ; Phosphodiesterase 4 Inhibitors - chemistry ; Phosphodiesterase 4 Inhibitors - metabolism ; Phosphodiesterase 4 Inhibitors - pharmacology ; Protein Conformation ; Protein kinase A ; Schizosaccharomyces - genetics ; Small Molecule Libraries - chemistry ; Small Molecule Libraries - metabolism ; Small Molecule Libraries - pharmacology ; Steroidogenesis ; Steroids - biosynthesis ; Studies ; Testosterone ; Yeast</subject><ispartof>PloS one, 2013-08, Vol.8 (8), p.e71279-e71279</ispartof><rights>COPYRIGHT 2013 Public Library of Science</rights><rights>2013 Demirbas et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License: https://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. 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BC8-15, BC8-15A, and BC8-15C, which are commercially available compounds, display distinct patterns of activity against PDE4, PDE8, PDE10A, and PDE11A, representing a chemical toolkit that could be used to examine the biological roles of these enzymes in cell culture systems.</description><subject>Adenosine</subject><subject>Animals</subject><subject>Assaying</subject><subject>Biological activity</subject><subject>Biology</subject><subject>Cell culture</subject><subject>Cell Line, Tumor</subject><subject>Cloning</subject><subject>Cyclic Nucleotide Phosphodiesterases, Type 4 - chemistry</subject><subject>Cyclic Nucleotide Phosphodiesterases, Type 4 - metabolism</subject><subject>Drug Evaluation, Preclinical - methods</subject><subject>Enzymes</subject><subject>Gene expression</subject><subject>Genetics</subject><subject>Glucose</subject><subject>High-Throughput Screening Assays</subject><subject>Humans</subject><subject>Inhibition</subject><subject>Inhibitors</subject><subject>Laboratory animals</subject><subject>Leydig cells</subject><subject>Leydig Cells - 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By replacing the Schizosaccharomyces pombe PDE gene with the murine PDE8A1 gene in strains lacking adenylyl cyclase, we generated strains whose protein kinase A (PKA)-stimulated growth in 5-fluoro orotic acid (5FOA) medium reflects PDE8 activity. From our previously-identified PDE4 and PDE7 inhibitors, we identified a PDE4/8 inhibitor that allowed us to optimize screening conditions. Of 222,711 compounds screened, ∼0.2% displayed composite Z scores of >20. Additional yeast-based assays using the most effective 367 compounds identified 30 candidates for further characterization. Among these, compound BC8-15 displayed the lowest IC₅₀ value for both PDE4 and PDE8 inhibition in in vitro enzyme assays. This compound also displays significant activity against PDE10A and PDE11A. BC8-15 elevates steroidogenesis in mouse Leydig cells as a single pharmacological agent. Assays using BC8-15 and two structural derivatives support a model in which PDE8 is a primary regulator of testosterone production by Leydig cells, with an additional role for PDE4 in this process. BC8-15, BC8-15A, and BC8-15C, which are commercially available compounds, display distinct patterns of activity against PDE4, PDE8, PDE10A, and PDE11A, representing a chemical toolkit that could be used to examine the biological roles of these enzymes in cell culture systems.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>23967182</pmid><doi>10.1371/journal.pone.0071279</doi><tpages>e71279</tpages><oa>free_for_read</oa></addata></record> |
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| identifier | ISSN: 1932-6203 |
| ispartof | PloS one, 2013-08, Vol.8 (8), p.e71279-e71279 |
| issn | 1932-6203 1932-6203 |
| language | eng |
| recordid | cdi_plos_journals_1430434352 |
| source | MEDLINE; DOAJ Directory of Open Access Journals; Public Library of Science (PLoS) Journals Open Access; EZB-FREE-00999 freely available EZB journals; PubMed Central; Free Full-Text Journals in Chemistry |
| subjects | Adenosine Animals Assaying Biological activity Biology Cell culture Cell Line, Tumor Cloning Cyclic Nucleotide Phosphodiesterases, Type 4 - chemistry Cyclic Nucleotide Phosphodiesterases, Type 4 - metabolism Drug Evaluation, Preclinical - methods Enzymes Gene expression Genetics Glucose High-Throughput Screening Assays Humans Inhibition Inhibitors Laboratory animals Leydig cells Leydig Cells - drug effects Leydig Cells - metabolism Lymphocytes Male Medical screening Mice Molecular Docking Simulation Orotic acid Pharmacology Phosphodiesterase Phosphodiesterase 4 Inhibitors - chemistry Phosphodiesterase 4 Inhibitors - metabolism Phosphodiesterase 4 Inhibitors - pharmacology Protein Conformation Protein kinase A Schizosaccharomyces - genetics Small Molecule Libraries - chemistry Small Molecule Libraries - metabolism Small Molecule Libraries - pharmacology Steroidogenesis Steroids - biosynthesis Studies Testosterone Yeast |
| title | A yeast-based chemical screen identifies a PDE inhibitor that elevates steroidogenesis in mouse Leydig cells via PDE8 and PDE4 inhibition |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-26T10%3A28%3A04IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=A%20yeast-based%20chemical%20screen%20identifies%20a%20PDE%20inhibitor%20that%20elevates%20steroidogenesis%20in%20mouse%20Leydig%20cells%20via%20PDE8%20and%20PDE4%20inhibition&rft.jtitle=PloS%20one&rft.au=Demirbas,%20Didem&rft.date=2013-08-14&rft.volume=8&rft.issue=8&rft.spage=e71279&rft.epage=e71279&rft.pages=e71279-e71279&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0071279&rft_dat=%3Cgale_plos_%3EA478288659%3C/gale_plos_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1430434352&rft_id=info:pmid/23967182&rft_galeid=A478288659&rft_doaj_id=oai_doaj_org_article_93923a8aea084547805499afdd9abfba&rfr_iscdi=true |