Targeting the PTPome in human disease

Tautz, Lutz; Pellecchia, Maurizio; Mustelin, Tomas

doi:10.1517/14728222.10.1.157

Cited by 100 publications

(64 citation statements)

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“…Protein tyrosine phosphatases are emerging as drug targets, but poor cell permeability of inhibitors has limited the development of drugs targeting these enzymes [Tautz L, et al (2006) Expert Opin Ther Targets 10:157-177]. Here we developed a method to monitor tyrosine phosphatase activity at the single-cell level and applied it to the identification of cell-permeable inhibitors.…”

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confidence: 99%

“…single-cell assay | peptide substrate P rotein tyrosine phosphatases (PTPs) are important regulators of signal transduction and are emerging as drug targets for common and debilitating human diseases, including cancer, diabetes, arthritis, and infectious diseases (1). The transmembrane PTP CD45, a critical regulator of signaling through the T-cell receptor (TCR), was one of the first PTPs to be considered as a drug target for treatment of human autoimmune diseases (2).…”

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High-throughput screen using a single-cell tyrosine phosphatase assay reveals biologically active inhibitors of tyrosine phosphatase CD45

Stanford

Panchal

Walker

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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Many cellular signaling events are regulated by tyrosine phosphorylation and mediated by the opposing actions of protein tyrosine kinases and phosphatases. Protein tyrosine phosphatases are emerging as drug targets, but poor cell permeability of inhibitors has limited the development of drugs targeting these enzymes [Tautz L, et al. (2006) Expert Opin Ther Targets 10:157-177]. Here we developed a method to monitor tyrosine phosphatase activity at the single-cell level and applied it to the identification of cell-permeable inhibitors. The method takes advantage of the fluorogenic properties of phosphorylated coumaryl amino propionic acid (pCAP), an analog of phosphotyrosine, which can be incorporated into peptides. Once delivered into cells, pCAP peptides were dephosphorylated by protein tyrosine phosphatases, and the resulting cell fluorescence could be monitored by flow cytometry and high-content imaging. The robustness and sensitivity of the assay was validated using peptides preferentially dephosphorylated by CD45 and T-cell tyrosine phosphatase and available inhibitors of these two enzymes. The assay was applied to high-throughput screening for inhibitors of CD45, an important target for autoimmunity and infectious diseases [Hermiston ML, et al. (2003) Annu Rev Immunol 21:107-137]. We identified four CD45 inhibitors that showed activity in T cells and macrophages. These results indicate that our assay can be applied to primary screening for inhibitors of CD45 and of other protein tyrosine phosphatases to increase the yield of biologically active inhibitors.single-cell assay | peptide substrate P rotein tyrosine phosphatases (PTPs) are important regulators of signal transduction and are emerging as drug targets for common and debilitating human diseases, including cancer, diabetes, arthritis, and infectious diseases (1). The transmembrane PTP CD45, a critical regulator of signaling through the T-cell receptor (TCR), was one of the first PTPs to be considered as a drug target for treatment of human autoimmune diseases (2). In addition, inhibitors of PTP-1B and SHP-2 currently are being sought for therapy of metabolic diseases and cancer, respectively (3-5). Unfortunately, development of small-molecule inhibitors of PTPs has been a challenging task. Poor cell permeability of PTP inhibitors has been a limitation difficult to overcome in later stages of drug development (5). Several methods to assess intracellular PTP activity are available (6-8), yet no assay has been able to capture intracellular PTP activity at the single-cell level. Cell-based PTP assays that work at the single-cell level and are suited to high-throughput screening might accelerate PTP inhibitor development.In this report we describe the development and optimization of a method to monitor PTP activity at the single-cell level by using cell-permeable fluorogenic peptides that, once delivered into cells, are dephosphorylated by PTPs. The dephosphorylation of these peptides by PTPs generates a signal that can be monitored by flow cytometry and h...

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confidence: 99%

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High-throughput screen using a single-cell tyrosine phosphatase assay reveals biologically active inhibitors of tyrosine phosphatase CD45

Stanford

Panchal

Walker

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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“…Our inhibitors contain a single salicylate linked to a furanyl moiety as phosphotyrosine mimic and a more variable group that could be exploited to achieve selectivity and higher affinity [8]. In fact, while very small differences can be seen in the phosphotyrosine binding pockets of tyrosine phosphatases, unique sub-pockets can be found in adjacent regions [3,8]. Therefore, it appears evident that by tailoring a second-site ligand, it should be possible to develop potent and selective inhibitors of therapeutically relevant protein tyrosine phosphatases.…”

Section: Introductionmentioning

confidence: 99%

Development of Molecular Probes for Second-Site Screening and Design of Protein Tyrosine Phosphatase Inhibitors

et al. 2007

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We report on the design, synthesis and evaluation of series of furanyl-salicyl-nitroxide derivatives, as effective chemical probes for second site screening against phosphotyrosine phosphatases (PTPs) using NMR-based techniques. The compounds have been tested against a panel of PTPs to asses their ability to inhibit a broad spectrum of these phosphatases. The utility of the derived compounds is illustrated with the phosphatase YopH, a bacterial toxin from Yersinia pestis. Novel chemical fragments were identified during a NMR-based screen for compounds that are capable to bind on the surface of YopH in regions adjacent the catalytic site in presence of the spin-labeled compounds. Our data demonstrate the value of the derived chemical probes for NMR-based second-site screening in PTPs.

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“…In contrast to the early hypothesis that PTP activity largely represents a basal ("housekeeping") counterforce to highly controlled protein-phosphorylation events (i.e., tyrosine kinase activity), it is now clear that PTP activity is also specific and tightly regulated, and that PTPs can exert either positive or negative effects on a signaling pathway [4][5][6]. Moreover, it is now beyond dispute that PTPs represent significant drug targets for a wide variety of clinically important pathologies [7][8][9][10]. Small-molecule inhibitors that can act specifically on individual PTPs would thus be important tools for both of these "worlds": understanding the basic-science roles of individual PTPs in complex signaling pathways, and validating PTPs as viable therapeutic targets [11,12].…”

Section: Introductionmentioning

confidence: 99%

Generation of inhibitor-sensitive protein tyrosine phosphatases via active-site mutations

Bishop

Zhang

Lone

2007

Methods

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Protein tyrosine phosphatases (PTPs) catalyze the dephosphorylation of phosphotyrosine, a central control element in mammalian signal transduction. Small-molecule inhibitors that are specific for each cellular PTP would be valuable tools in dissecting phosphorylation networks and for validating PTPs as therapeutic targets. However, the common architecture of PTP active sites impedes the discovery of selective PTP inhibitors. Our laboratory has recently used enzyme/inhibitor-interface engineering to generate selective PTP inhibitors. The crux of the strategy resides in the design of "inhibitor-sensitized" PTPs through protein engineering of a novel binding pocket in the target PTP. "Allele-specific" inhibitors that selectively target the sensitized PTP can be synthesized by modifying broad-specificity inhibitors with bulky chemical groups that are incompatible with wild-type PTP active sites; alternatively, specific inhibitors that serendipitously recognize the sensitized PTP's nonnatural pocket may be discovered from panels of "non-rationally" designed compounds. In this review, we describe the current state of the PTP-sensitization strategy, with emphases on the methodology of identifying PTP-sensitizing mutations and synthesizing the compounds that have been found to target PTPs in an allele-specific manner. Moreover, we discuss the scope of PTP sensitization in regard to the potential application of the approach across the family of classical PTPs.

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Targeting the PTPome in human disease

Cited by 100 publications

References 177 publications

High-throughput screen using a single-cell tyrosine phosphatase assay reveals biologically active inhibitors of tyrosine phosphatase CD45

High-throughput screen using a single-cell tyrosine phosphatase assay reveals biologically active inhibitors of tyrosine phosphatase CD45

Development of Molecular Probes for Second-Site Screening and Design of Protein Tyrosine Phosphatase Inhibitors

Generation of inhibitor-sensitive protein tyrosine phosphatases via active-site mutations

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