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Cdc25B Dual-specificity phosphatase inhibitors identified in a high-throughput screen of the NIH compound library

Johnston, PA and Foster, CA and Tierno, MB and Shun, TY and Shinde, SN and Paquette, WD and Brummond, KM and Wipf, P and Lazo, JS (2009) Cdc25B Dual-specificity phosphatase inhibitors identified in a high-throughput screen of the NIH compound library. Assay and Drug Development Technologies, 7 (3). 250 - 265. ISSN 1540-658X

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Abstract

The University of Pittsburgh Molecular Library Screening Center (Pittsburgh, PA) conducted a screen with the National Institutes of Health compound library for inhibitors of in vitro cell division cycle 25 protein (Cdc25) B activity during the pilot phase of the Molecular Library Screening Center Network. Seventy-nine (0.12%) of the 65,239 compounds screened at 10 μM met the active criterion of ≥50% inhibition of Cdc25B activity, and 25 (31.6%) of these were confirmed as Cdc25B inhibitors with 50% inhibitory concentration (IC ) values <50 μM. Thirteen of the Cdc25B inhibitors were represented by singleton chemical structures, and 12 were divided among four clusters of related structures. Thirteen (52%) of the Cdc25B inhibitor hits were quinone-based structures. The Cdc25B inhibitors were further characterized in a series of in vitro secondary assays to confirm their activity, to determine their phosphatase selectivity against two other dual-specificity phosphatases, mitogen-activated protein kinase phosphatase (MKP)-1 and MKP-3, and to examine if the mechanism of Cdc25B inhibition involved oxidation and inactivation. Nine Cdc25B inhibitors did not appear to affect Cdc25B through a mechanism involving oxidation because they did not generate detectable amounts of H O in the presence of dithiothreitol, and their Cdc25B IC values were not significantly affected by exchanging the dithiothreitol for β-mercaptoethanol or reduced glutathione or by adding catalase to the assay. Six of the nonoxidative hits were selective for Cdc25B inhibition versus MKP-1 and MKP-3, but only the two bisfuran-containing hits, PubChem substance identifiers 4258795 and 4260465, significantly inhibited the growth of human MBA-MD-435 breast and PC-3 prostate cancer cell lines. To confirm the structure and biological activity of 4260465, the compound was resynthesized along with two analogs. Neither of the substitutions to the two analogs was tolerated, and only the resynthesized hit 26683752 inhibited Cdc25B activity in vitro (IC = 13.83 ± 1.0 μM) and significantly inhibited the growth of the MBA-MD-435 breast and PC-3 prostate cancer cell lines (IC = 20.16 ± 2.0 μM and 24.87 ± 2.25 μM, respectively). The two bis-furan-containing hits identified in the screen represent novel nonoxidative Cdc25B inhibitor chemotypes that block tumor cell proliferation. The availability of non-redox active Cdc25B inhibitors should provide valuable tools to explore the inhibition of the Cdc25 phosphatases as potential mono- or combination therapies for cancer. © 2009, Mary Ann Liebert, Inc. 50 2 2 50 50 50


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Details

Item Type: Article
Status: Published
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Johnston, PApaj18@pitt.eduPAJ180000-0001-5815-3091
Foster, CA
Tierno, MB
Shun, TY
Shinde, SN
Paquette, WD
Brummond, KMkbrummon@pitt.eduKBRUMMON
Wipf, Ppwipf@pitt.eduPWIPF
Lazo, JS
Date: 1 June 2009
Date Type: Publication
Journal or Publication Title: Assay and Drug Development Technologies
Volume: 7
Number: 3
Page Range: 250 - 265
DOI or Unique Handle: 10.1089/adt.2008.186
Schools and Programs: Dietrich School of Arts and Sciences > Chemistry
Refereed: Yes
ISSN: 1540-658X
MeSH Headings: Antineoplastic Agents--chemistry; Antineoplastic Agents--pharmacology; Cell Line, Tumor; Drug Evaluation, Preclinical; Dual Specificity Phosphatase 1--antagonists & inhibitors; Dual Specificity Phosphatase 1--biosynthesis; Dual Specificity Phosphatase 1--isolation & purification; Dual Specificity Phosphatase 6--antagonists & inhibitors; Dual Specificity Phosphatase 6--biosynthesis; Dual Specificity Phosphatase 6--isolation & purification; Enzyme Inhibitors--chemistry; Enzyme Inhibitors--pharmacology; Female; Humans; Hydrogen Peroxide--chemistry; Indicators and Reagents; Male; Mitogen-Activated Protein Kinase Phosphatases--antagonists & inhibitors; Mitogen-Activated Protein Kinase Phosphatases--biosynthesis; Mitogen-Activated Protein Kinase Phosphatases--isolation & purification; National Institutes of Health (U.S.); Oxidation-Reduction; Small Molecule Libraries; United States; cdc25 Phosphatases--antagonists & inhibitors
Other ID: NLM PMC2956648
PubMed Central ID: PMC2956648
PubMed ID: 19530895
Date Deposited: 18 Jun 2013 20:44
Last Modified: 30 Mar 2021 12:55
URI: http://d-scholarship.pitt.edu/id/eprint/18955

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