Differential selectivity of JAK2 inhibitors in enzymatic and cellular settings
Small molecule inhibitors of Janus kinase (JAK) family members (JAK1, JAK2, JAK3, and Tyk2) are currently being pursued as potential new modes of therapy for a variety of diseases, including the inhibition of JAK2 for the treatment of myeloproliferative disorders. Selective inhibition within the JAK...
Gespeichert in:
| Veröffentlicht in: | Experimental hematology 2013-05, Vol.41 (5), p.491-500 |
|---|---|
| Hauptverfasser: | , , , , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 500 |
|---|---|
| container_issue | 5 |
| container_start_page | 491 |
| container_title | Experimental hematology |
| container_volume | 41 |
| creator | Yu, Violeta Pistillo, Jeanne Archibeque, Ivonne Han Lee, Josie Sun, Bee-Chun Schenkel, Laurie B Geuns-Meyer, Stephanie Liu, Liqin Emkey, Renee |
| description | Small molecule inhibitors of Janus kinase (JAK) family members (JAK1, JAK2, JAK3, and Tyk2) are currently being pursued as potential new modes of therapy for a variety of diseases, including the inhibition of JAK2 for the treatment of myeloproliferative disorders. Selective inhibition within the JAK family can be beneficial in avoiding undesirable side effects (e.g., immunosuppression) caused by parallel inhibition of other JAK members. In an effort to design an assay paradigm for the development of JAK2 selective inhibitors, we investigated whether compound selectivity differed between cellular and purified enzyme environments. A set of JAK2 inhibitors was tested in a high-throughput JAK family cell assay suite and in corresponding purified enzyme assays. The high-throughput JAK cell assay suite comprises Ba/F3 cells individually expressing translocated ETS leukemia (TEL) fusions of each JAK family member (TEL-JAK Ba/F3) and an AlphaScreen phosphorylated-STAT5 (pSTAT5) immunoassay. Compound potencies from the TEL-JAK Ba/F3 pSTAT5 assays were similar to those determined in downstream cell proliferation measurements and more physiologically relevant cytokine-induced pSTAT5 PBMC assays. However, compound selectivity data between cell and purified enzyme assays were discrepant because of different potency shifts between cell and purified enzyme values for each JAK family member. For any JAK small molecule development program, our results suggest that relying solely on enzyme potency and selectivity data may be misleading. Adopting the high-throughput TEL-JAK Ba/F3 pSTAT5 cell assay suite in lead development paradigms should provide a more meaningful understanding of selectivity and facilitate the development of more selective JAK inhibitors. |
| doi_str_mv | 10.1016/j.exphem.2013.01.005 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_1347788545</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>S0301472X13000064</els_id><sourcerecordid>1347788545</sourcerecordid><originalsourceid>FETCH-LOGICAL-c463t-1368c4f2aa782ffecd586e724b8d6dab48220fbc5970d9becc908d57e6fa68e63</originalsourceid><addsrcrecordid>eNqFkUuP1DAQhC0EYoeFf4BQjlwS_IrjXJBWy5sVHACJm-XYHbaHxBlsZ8Xw63E0CwcunNqHqq72V4Q8ZrRhlKln-wZ-Hq5hbjhloqGsobS9Q3ZMd6Lmou_vkh0VlNWy41_PyIOU9rQo2p7eJ2dcCFlcakc-vMBxhAgho52qBBO4jDeYj9UyVu8u3vMKwzUOmJeYyrOC8Os424yussFXDqZpnWwsxpwxfEsPyb3RTgke3c5z8uXVy8-Xb-qrj6_fXl5c1U4qkesSrZ0cubWd5iXf-VYr6LgctFfeDlJzTsfBtX1HfT-Acz3Vvu1AjVZpUOKcPD3tPcTlxwopmxnTdo0NsKzJMCG7TutWtkUqT1IXl5QijOYQcbbxaBg1G0mzNyeSZiNpKDOFU7E9uU1Yhxn8X9MfdEXw_CSA8s8bhGiSQwgOPMYC0fgF_5fw7wI3YUBnp-9whLRf1hgKQ8NM4oaaT1ubW5lMlCKpkuI3UxecRA</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1347788545</pqid></control><display><type>article</type><title>Differential selectivity of JAK2 inhibitors in enzymatic and cellular settings</title><source>MEDLINE</source><source>Elsevier ScienceDirect Journals</source><creator>Yu, Violeta ; Pistillo, Jeanne ; Archibeque, Ivonne ; Han Lee, Josie ; Sun, Bee-Chun ; Schenkel, Laurie B ; Geuns-Meyer, Stephanie ; Liu, Liqin ; Emkey, Renee</creator><creatorcontrib>Yu, Violeta ; Pistillo, Jeanne ; Archibeque, Ivonne ; Han Lee, Josie ; Sun, Bee-Chun ; Schenkel, Laurie B ; Geuns-Meyer, Stephanie ; Liu, Liqin ; Emkey, Renee</creatorcontrib><description>Small molecule inhibitors of Janus kinase (JAK) family members (JAK1, JAK2, JAK3, and Tyk2) are currently being pursued as potential new modes of therapy for a variety of diseases, including the inhibition of JAK2 for the treatment of myeloproliferative disorders. Selective inhibition within the JAK family can be beneficial in avoiding undesirable side effects (e.g., immunosuppression) caused by parallel inhibition of other JAK members. In an effort to design an assay paradigm for the development of JAK2 selective inhibitors, we investigated whether compound selectivity differed between cellular and purified enzyme environments. A set of JAK2 inhibitors was tested in a high-throughput JAK family cell assay suite and in corresponding purified enzyme assays. The high-throughput JAK cell assay suite comprises Ba/F3 cells individually expressing translocated ETS leukemia (TEL) fusions of each JAK family member (TEL-JAK Ba/F3) and an AlphaScreen phosphorylated-STAT5 (pSTAT5) immunoassay. Compound potencies from the TEL-JAK Ba/F3 pSTAT5 assays were similar to those determined in downstream cell proliferation measurements and more physiologically relevant cytokine-induced pSTAT5 PBMC assays. However, compound selectivity data between cell and purified enzyme assays were discrepant because of different potency shifts between cell and purified enzyme values for each JAK family member. For any JAK small molecule development program, our results suggest that relying solely on enzyme potency and selectivity data may be misleading. Adopting the high-throughput TEL-JAK Ba/F3 pSTAT5 cell assay suite in lead development paradigms should provide a more meaningful understanding of selectivity and facilitate the development of more selective JAK inhibitors.</description><identifier>ISSN: 0301-472X</identifier><identifier>EISSN: 1873-2399</identifier><identifier>DOI: 10.1016/j.exphem.2013.01.005</identifier><identifier>PMID: 23340136</identifier><language>eng</language><publisher>Netherlands: Elsevier Inc</publisher><subject>Advanced Basic Science ; Animals ; Cell Line ; Cell Proliferation - drug effects ; Cells, Cultured ; Dose-Response Relationship, Drug ; Hematology, Oncology and Palliative Medicine ; High-Throughput Screening Assays - methods ; Humans ; Janus Kinase 2 - antagonists & inhibitors ; Janus Kinase 2 - genetics ; Janus Kinase 2 - metabolism ; Leukocytes, Mononuclear - drug effects ; Leukocytes, Mononuclear - metabolism ; Oncogene Proteins, Fusion - antagonists & inhibitors ; Oncogene Proteins, Fusion - genetics ; Oncogene Proteins, Fusion - metabolism ; Phosphorylation - drug effects ; Precursor Cells, B-Lymphoid - drug effects ; Precursor Cells, B-Lymphoid - metabolism ; Protein Kinase Inhibitors - pharmacology ; Reproducibility of Results ; STAT5 Transcription Factor - metabolism</subject><ispartof>Experimental hematology, 2013-05, Vol.41 (5), p.491-500</ispartof><rights>ISEH - Society for Hematology and Stem Cells</rights><rights>2013 ISEH - Society for Hematology and Stem Cells</rights><rights>Copyright © 2013 ISEH - Society for Hematology and Stem Cells. Published by Elsevier Inc. All rights reserved.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c463t-1368c4f2aa782ffecd586e724b8d6dab48220fbc5970d9becc908d57e6fa68e63</citedby><cites>FETCH-LOGICAL-c463t-1368c4f2aa782ffecd586e724b8d6dab48220fbc5970d9becc908d57e6fa68e63</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0301472X13000064$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/23340136$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Yu, Violeta</creatorcontrib><creatorcontrib>Pistillo, Jeanne</creatorcontrib><creatorcontrib>Archibeque, Ivonne</creatorcontrib><creatorcontrib>Han Lee, Josie</creatorcontrib><creatorcontrib>Sun, Bee-Chun</creatorcontrib><creatorcontrib>Schenkel, Laurie B</creatorcontrib><creatorcontrib>Geuns-Meyer, Stephanie</creatorcontrib><creatorcontrib>Liu, Liqin</creatorcontrib><creatorcontrib>Emkey, Renee</creatorcontrib><title>Differential selectivity of JAK2 inhibitors in enzymatic and cellular settings</title><title>Experimental hematology</title><addtitle>Exp Hematol</addtitle><description>Small molecule inhibitors of Janus kinase (JAK) family members (JAK1, JAK2, JAK3, and Tyk2) are currently being pursued as potential new modes of therapy for a variety of diseases, including the inhibition of JAK2 for the treatment of myeloproliferative disorders. Selective inhibition within the JAK family can be beneficial in avoiding undesirable side effects (e.g., immunosuppression) caused by parallel inhibition of other JAK members. In an effort to design an assay paradigm for the development of JAK2 selective inhibitors, we investigated whether compound selectivity differed between cellular and purified enzyme environments. A set of JAK2 inhibitors was tested in a high-throughput JAK family cell assay suite and in corresponding purified enzyme assays. The high-throughput JAK cell assay suite comprises Ba/F3 cells individually expressing translocated ETS leukemia (TEL) fusions of each JAK family member (TEL-JAK Ba/F3) and an AlphaScreen phosphorylated-STAT5 (pSTAT5) immunoassay. Compound potencies from the TEL-JAK Ba/F3 pSTAT5 assays were similar to those determined in downstream cell proliferation measurements and more physiologically relevant cytokine-induced pSTAT5 PBMC assays. However, compound selectivity data between cell and purified enzyme assays were discrepant because of different potency shifts between cell and purified enzyme values for each JAK family member. For any JAK small molecule development program, our results suggest that relying solely on enzyme potency and selectivity data may be misleading. Adopting the high-throughput TEL-JAK Ba/F3 pSTAT5 cell assay suite in lead development paradigms should provide a more meaningful understanding of selectivity and facilitate the development of more selective JAK inhibitors.</description><subject>Advanced Basic Science</subject><subject>Animals</subject><subject>Cell Line</subject><subject>Cell Proliferation - drug effects</subject><subject>Cells, Cultured</subject><subject>Dose-Response Relationship, Drug</subject><subject>Hematology, Oncology and Palliative Medicine</subject><subject>High-Throughput Screening Assays - methods</subject><subject>Humans</subject><subject>Janus Kinase 2 - antagonists & inhibitors</subject><subject>Janus Kinase 2 - genetics</subject><subject>Janus Kinase 2 - metabolism</subject><subject>Leukocytes, Mononuclear - drug effects</subject><subject>Leukocytes, Mononuclear - metabolism</subject><subject>Oncogene Proteins, Fusion - antagonists & inhibitors</subject><subject>Oncogene Proteins, Fusion - genetics</subject><subject>Oncogene Proteins, Fusion - metabolism</subject><subject>Phosphorylation - drug effects</subject><subject>Precursor Cells, B-Lymphoid - drug effects</subject><subject>Precursor Cells, B-Lymphoid - metabolism</subject><subject>Protein Kinase Inhibitors - pharmacology</subject><subject>Reproducibility of Results</subject><subject>STAT5 Transcription Factor - metabolism</subject><issn>0301-472X</issn><issn>1873-2399</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2013</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkUuP1DAQhC0EYoeFf4BQjlwS_IrjXJBWy5sVHACJm-XYHbaHxBlsZ8Xw63E0CwcunNqHqq72V4Q8ZrRhlKln-wZ-Hq5hbjhloqGsobS9Q3ZMd6Lmou_vkh0VlNWy41_PyIOU9rQo2p7eJ2dcCFlcakc-vMBxhAgho52qBBO4jDeYj9UyVu8u3vMKwzUOmJeYyrOC8Os424yussFXDqZpnWwsxpwxfEsPyb3RTgke3c5z8uXVy8-Xb-qrj6_fXl5c1U4qkesSrZ0cubWd5iXf-VYr6LgctFfeDlJzTsfBtX1HfT-Acz3Vvu1AjVZpUOKcPD3tPcTlxwopmxnTdo0NsKzJMCG7TutWtkUqT1IXl5QijOYQcbbxaBg1G0mzNyeSZiNpKDOFU7E9uU1Yhxn8X9MfdEXw_CSA8s8bhGiSQwgOPMYC0fgF_5fw7wI3YUBnp-9whLRf1hgKQ8NM4oaaT1ubW5lMlCKpkuI3UxecRA</recordid><startdate>20130501</startdate><enddate>20130501</enddate><creator>Yu, Violeta</creator><creator>Pistillo, Jeanne</creator><creator>Archibeque, Ivonne</creator><creator>Han Lee, Josie</creator><creator>Sun, Bee-Chun</creator><creator>Schenkel, Laurie B</creator><creator>Geuns-Meyer, Stephanie</creator><creator>Liu, Liqin</creator><creator>Emkey, Renee</creator><general>Elsevier Inc</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>20130501</creationdate><title>Differential selectivity of JAK2 inhibitors in enzymatic and cellular settings</title><author>Yu, Violeta ; Pistillo, Jeanne ; Archibeque, Ivonne ; Han Lee, Josie ; Sun, Bee-Chun ; Schenkel, Laurie B ; Geuns-Meyer, Stephanie ; Liu, Liqin ; Emkey, Renee</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c463t-1368c4f2aa782ffecd586e724b8d6dab48220fbc5970d9becc908d57e6fa68e63</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2013</creationdate><topic>Advanced Basic Science</topic><topic>Animals</topic><topic>Cell Line</topic><topic>Cell Proliferation - drug effects</topic><topic>Cells, Cultured</topic><topic>Dose-Response Relationship, Drug</topic><topic>Hematology, Oncology and Palliative Medicine</topic><topic>High-Throughput Screening Assays - methods</topic><topic>Humans</topic><topic>Janus Kinase 2 - antagonists & inhibitors</topic><topic>Janus Kinase 2 - genetics</topic><topic>Janus Kinase 2 - metabolism</topic><topic>Leukocytes, Mononuclear - drug effects</topic><topic>Leukocytes, Mononuclear - metabolism</topic><topic>Oncogene Proteins, Fusion - antagonists & inhibitors</topic><topic>Oncogene Proteins, Fusion - genetics</topic><topic>Oncogene Proteins, Fusion - metabolism</topic><topic>Phosphorylation - drug effects</topic><topic>Precursor Cells, B-Lymphoid - drug effects</topic><topic>Precursor Cells, B-Lymphoid - metabolism</topic><topic>Protein Kinase Inhibitors - pharmacology</topic><topic>Reproducibility of Results</topic><topic>STAT5 Transcription Factor - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Yu, Violeta</creatorcontrib><creatorcontrib>Pistillo, Jeanne</creatorcontrib><creatorcontrib>Archibeque, Ivonne</creatorcontrib><creatorcontrib>Han Lee, Josie</creatorcontrib><creatorcontrib>Sun, Bee-Chun</creatorcontrib><creatorcontrib>Schenkel, Laurie B</creatorcontrib><creatorcontrib>Geuns-Meyer, Stephanie</creatorcontrib><creatorcontrib>Liu, Liqin</creatorcontrib><creatorcontrib>Emkey, Renee</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Experimental hematology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Yu, Violeta</au><au>Pistillo, Jeanne</au><au>Archibeque, Ivonne</au><au>Han Lee, Josie</au><au>Sun, Bee-Chun</au><au>Schenkel, Laurie B</au><au>Geuns-Meyer, Stephanie</au><au>Liu, Liqin</au><au>Emkey, Renee</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Differential selectivity of JAK2 inhibitors in enzymatic and cellular settings</atitle><jtitle>Experimental hematology</jtitle><addtitle>Exp Hematol</addtitle><date>2013-05-01</date><risdate>2013</risdate><volume>41</volume><issue>5</issue><spage>491</spage><epage>500</epage><pages>491-500</pages><issn>0301-472X</issn><eissn>1873-2399</eissn><abstract>Small molecule inhibitors of Janus kinase (JAK) family members (JAK1, JAK2, JAK3, and Tyk2) are currently being pursued as potential new modes of therapy for a variety of diseases, including the inhibition of JAK2 for the treatment of myeloproliferative disorders. Selective inhibition within the JAK family can be beneficial in avoiding undesirable side effects (e.g., immunosuppression) caused by parallel inhibition of other JAK members. In an effort to design an assay paradigm for the development of JAK2 selective inhibitors, we investigated whether compound selectivity differed between cellular and purified enzyme environments. A set of JAK2 inhibitors was tested in a high-throughput JAK family cell assay suite and in corresponding purified enzyme assays. The high-throughput JAK cell assay suite comprises Ba/F3 cells individually expressing translocated ETS leukemia (TEL) fusions of each JAK family member (TEL-JAK Ba/F3) and an AlphaScreen phosphorylated-STAT5 (pSTAT5) immunoassay. Compound potencies from the TEL-JAK Ba/F3 pSTAT5 assays were similar to those determined in downstream cell proliferation measurements and more physiologically relevant cytokine-induced pSTAT5 PBMC assays. However, compound selectivity data between cell and purified enzyme assays were discrepant because of different potency shifts between cell and purified enzyme values for each JAK family member. For any JAK small molecule development program, our results suggest that relying solely on enzyme potency and selectivity data may be misleading. Adopting the high-throughput TEL-JAK Ba/F3 pSTAT5 cell assay suite in lead development paradigms should provide a more meaningful understanding of selectivity and facilitate the development of more selective JAK inhibitors.</abstract><cop>Netherlands</cop><pub>Elsevier Inc</pub><pmid>23340136</pmid><doi>10.1016/j.exphem.2013.01.005</doi><tpages>10</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0301-472X |
| ispartof | Experimental hematology, 2013-05, Vol.41 (5), p.491-500 |
| issn | 0301-472X 1873-2399 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_1347788545 |
| source | MEDLINE; Elsevier ScienceDirect Journals |
| subjects | Advanced Basic Science Animals Cell Line Cell Proliferation - drug effects Cells, Cultured Dose-Response Relationship, Drug Hematology, Oncology and Palliative Medicine High-Throughput Screening Assays - methods Humans Janus Kinase 2 - antagonists & inhibitors Janus Kinase 2 - genetics Janus Kinase 2 - metabolism Leukocytes, Mononuclear - drug effects Leukocytes, Mononuclear - metabolism Oncogene Proteins, Fusion - antagonists & inhibitors Oncogene Proteins, Fusion - genetics Oncogene Proteins, Fusion - metabolism Phosphorylation - drug effects Precursor Cells, B-Lymphoid - drug effects Precursor Cells, B-Lymphoid - metabolism Protein Kinase Inhibitors - pharmacology Reproducibility of Results STAT5 Transcription Factor - metabolism |
| title | Differential selectivity of JAK2 inhibitors in enzymatic and cellular settings |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-08T22%3A02%3A02IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Differential%20selectivity%20of%20JAK2%20inhibitors%20in%20enzymatic%20and%20cellular%20settings&rft.jtitle=Experimental%20hematology&rft.au=Yu,%20Violeta&rft.date=2013-05-01&rft.volume=41&rft.issue=5&rft.spage=491&rft.epage=500&rft.pages=491-500&rft.issn=0301-472X&rft.eissn=1873-2399&rft_id=info:doi/10.1016/j.exphem.2013.01.005&rft_dat=%3Cproquest_cross%3E1347788545%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1347788545&rft_id=info:pmid/23340136&rft_els_id=S0301472X13000064&rfr_iscdi=true |