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5 results on '"Carola Seyffarth"'

1. An unconventional gatekeeper mutation sensitizes inositol hexakisphosphate kinases to an allosteric inhibitor

Author

Tim Aguirre, Gillian L Dornan, Sarah Hostachy, Martin Neuenschwander, Carola Seyffarth, Volker Haucke, Anja Schütz, Jens Peter von Kries, and Dorothea Fiedler

Subjects
kinase, inositol phosphate, allosteric inhibitor, analog-sensitive, Medicine, Science, Biology (General), QH301-705.5
Abstract

Inositol hexakisphosphate kinases (IP6Ks) are emerging as relevant pharmacological targets because a multitude of disease-related phenotypes has been associated with their function. While the development of potent IP6K inhibitors is gaining momentum, a pharmacological tool to distinguish the mammalian isozymes is still lacking. Here, we implemented an analog-sensitive approach for IP6Ks and performed a high-throughput screen to identify suitable lead compounds. The most promising hit, FMP-201300, exhibited high potency and selectivity toward the unique valine gatekeeper mutants of IP6K1 and IP6K2, compared to the respective wild-type (WT) kinases. Biochemical validation experiments revealed an allosteric mechanism of action that was corroborated by hydrogen deuterium exchange mass spectrometry measurements. The latter analysis suggested that displacement of the αC helix, caused by the gatekeeper mutation, facilitates the binding of FMP-201300 to an allosteric pocket adjacent to the ATP-binding site. FMP-201300 therefore serves as a valuable springboard for the further development of compounds that can selectively target the three mammalian IP6Ks; either as analog-sensitive kinase inhibitors or as an allosteric lead compound for the WT kinases.

Published
2023
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2. From Pyrazolones to Azaindoles: Evolution of Active-Site SHP2 Inhibitors Based on Scaffold Hopping and Bioisosteric Replacement

Author

Sandra Miksche, Nandor Ziebart, Yelena Mostinski, Judith de Schryver, Sylvia Oestreich, Christoph Rademacher, Yuliya Kolomeets, Martin Neuenschwander, Walter Birchmeier, Carola Seyffarth, Silke Radetzki, María P. López-Alberca, Guus J.J.E. Heynen, Jens Peter von Kries, Marc Nazaré, Elena Shanina, Yvette Roske, Jerome Paul, David Schaller, Andrea Volkamer, and Udo Heinemann

Subjects
MAPK/ERK pathway, Indoles, MAP Kinase Signaling System, Antineoplastic Agents, Protein Tyrosine Phosphatase, Non-Receptor Type 11, Protein tyrosine phosphatase, Molecular Dynamics Simulation, medicine.disease_cause, Metastasis, Structure-Activity Relationship, Catalytic Domain, Cell Line, Tumor, Drug Discovery, medicine, Humans, Enzyme Inhibitors, Pyrazolones, Cell Proliferation, Molecular Structure, Chemistry, Cell growth, Cancer, medicine.disease, Molecular Docking Simulation, Cancer research, Molecular Medicine, Drug Screening Assays, Antitumor, Signal transduction, Carcinogenesis, Protein Binding
Abstract

The tyrosine phosphatase SHP2 controls the activity of pivotal signaling pathways, including MAPK, JAK-STAT, and PI3K-Akt. Aberrant SHP2 activity leads to uncontrolled cell proliferation, tumorigenesis, and metastasis. SHP2 signaling was recently linked to drug resistance against cancer medications such as MEK and BRAF inhibitors. In this work, we present the development of a novel class of azaindole SHP2 inhibitors. We applied scaffold hopping and bioisosteric replacement concepts to eliminate unwanted structural motifs and to improve the inhibitor characteristics of the previously reported pyrazolone SHP2 inhibitors. The most potent azaindole 45 inhibits SHP2 with an IC50 = 0.031 μM in an enzymatic assay and with an IC50 = 2.6 μM in human pancreas cells (HPAF-II). Evaluation in a series of cellular assays for metastasis and drug resistance demonstrated efficient SHP2 blockade. Finally, 45 inhibited proliferation of two cancer cell lines that are resistant to cancer drugs and diminished ERK signaling.

Published
2020

5. Identification of a new membrane-permeable inhibitor against inositol-1,4,5-trisphosphate-3-kinase A

Author

Dominik Schröder, Martin Neuenschwander, Jens Peter von Kries, Christoph Rehbach, Carola Seyffarth, and Sabine Windhorst

Subjects
Cell Membrane Permeability, Membrane permeability, Kinase, Biophysics, Drug Evaluation, Preclinical, chemistry.chemical_element, Cell Biology, Calcium, Biology, Biochemistry, Small molecule, Cell biology, High-Throughput Screening Assays, Small Molecule Libraries, chemistry.chemical_compound, Phosphotransferases (Alcohol Group Acceptor), chemistry, Cell Line, Tumor, Phosphorylation, Humans, Ectopic expression, Inositol, Molecular Biology, Protein Kinase Inhibitors, Actin
Abstract

Ectopic expression of the neuron-specific inositol-1,4,5-trisphosphate-3-kinase A (ITPKA) in lung cancer cells increases their metastatic potential because the protein exhibits two actin regulating activities; it bundles actin filaments and regulates inositol-1,4,5-trisphosphate (InsP3)-mediated calcium signals by phosphorylating InsP3. Thus, in order to inhibit the metastasis-promoting activity of ITPKA, both its actin bundling and its InsP3kinase activity has to be blocked. In this study, we performed a high throughput screen in order to identify specific and membrane-permeable substances against the InsP3kinase activity. Among 341,44 small molecules, 237 compounds (0.7%) were identified as potential InsP3kinase inhibitors. After determination of IC50-values, the three compounds with highest specificity and highest hydrophobicity (EPPC-3, BAMB-4, MEPTT-3) were further characterized. Only BAMB-4 was nearly completely taken up by H1299 cells and remained stable after cellular uptake, thus exhibiting a robust stability and a high membrane permeability. Determination of the inhibitor type revealed that BAMB-4 belongs to the group of mixed type inhibitors. Taken together, for the first time we identified a highly membrane-permeable inhibitor against the InsP3kinase activity of ITPKA providing the possibility to partly inhibit the metastasis-promoting effect of ITPKA in lung tumor cells.

Published
2013

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