Skepinone-L is a selective p38 mitogen-activated protein kinase inhibitor

Koeberle, Solveigh C.; Romir, Johannes; Fischer, Stefan; Koeberle, Andreas; Schattel, Verena; Albrecht, Wolfgang; Grütter, Christian; Werz, Oliver; Rauh, Daniel; Stehle, Thilo; Laufer, Stefan

doi:10.1038/nchembio.761

Cited by 110 publications

(154 citation statements)

References 25 publications

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“…However, lack of high potency and selectivity inhibitors in vitro and especially in vivo has hampered investigation of p38 MAPK signaling pathways in platelet activation and pathological thrombus formation. The novel dibenzosuberone-type p38 MAPK inhibitor Skepinone-L shows outstanding selectivity and high in vivo potency [25]. …”

Section: Discussionmentioning

confidence: 99%

“…Skepinone-L was generated by S. Laufer (University of Tübingen, Germany) as described previously [25]. …”

Section: Methodsmentioning

confidence: 99%

“…Recently, Skepinone-L, a highly selective inhibitor of p38 MAPK has been developed. Skepinone-L is, to our knowledge, the first ATP-competitive p38 MAPK inhibitor with excellent in vivo efficacy and selectivity [25,27]. At a concentration of 1 µM, Skepinone-L did not bind any of the tested kinases in two different selectivity screens except p38 MAPK [25].…”

Section: Introductionmentioning

confidence: 99%

“…The majority of p38 MAPK inhibitors are only moderately potent in physiologically relevant whole blood studies [24] and require high dosing in vivo to achieve effective plasma concentrations [25]. Most p38 inhibitors including SB203580 and SB202190 are effective via their ATP-competitive binding mode and display poor selectivity because the ATP-binding site is highly conserved among a variety of proteins [25,26]. Recently, Skepinone-L, a highly selective inhibitor of p38 MAPK has been developed.…”

Section: Introductionmentioning

confidence: 99%

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Skepinone-L, a Novel Potent and Highly Selective Inhibitor of p38 MAP Kinase, Effectively Impairs Platelet Activation and Thrombus Formation

Borst

Walker²,

Münzer³

et al. 2013

Cell Physiol Biochem

1,339

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Background/Aims: Platelets are critically important for primary haemostasis and the major players in thrombotic vascular occlusion. Platelets are activated by agonists, such as thrombin and collagen-related peptide as well as second-wave mediators including thromboxane A₂ via different intracellular signaling pathways resulting in degranulation, aggregation and thrombus formation. Platelet activation is paralleled by phosphorylation and activation of p38 MAPK. The limited specificity of hitherto known p38 MAPK inhibitors precluded safe conclusions on the precise role of p38 MAPK in the regulation of platelet function. The present study examined the impact of Skepinone-L, a novel and highly selective inhibitor of p38 mitogen-activated protein kinase (p38 MAPK), on platelet activation and thrombus formation. Methods: Experiments were performed in freshly isolated human platelets. Protein phosphorylation was quantified by Western blotting, thromboxane B₂ synthesis by enzyme immunoassay, ATP release by ChronoLume luciferin assay, cytosolic Ca²⁺ concentration by Fura-2 fluorescence-measurements, platelet aggregation by a light transmissions measurement and in vitro thrombus formation by a flow chamber. Results: Skepinone-L (1 μM) virtually abrogated the phosphorylation of platelet p38 MAPK substrate Hsp27 following stimulation with CRP (1 μg/ml), thrombin (5 mU/ml) or thromboxane A₂ analogue U-46619 (1 μM). Furthermore, Skepinone-L significantly blunted activation-dependent platelet secretion and aggregation following threshold concentrations of CRP, thrombin and thromboxane A₂ analogue U-46619. Skepinone-L did not impair platelet Ca²⁺ signaling but prevented agonist-induced thromboxane A₂ synthesis through abrogation of p38 MAPK-dependent phosphorylation of platelet cytosolic phospholipase A₂ (cPLA₂). Skepinone-L further markedly blunted thrombus formation under low (500-s) and high (1700-s) arterial shear rates. Conclusions: The present study discloses a powerful inhibiting effect of p38 MAPK-blocker Skepinone-L on platelet activation and thrombus formation.

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Section: Discussionmentioning

confidence: 99%

“…Skepinone-L was generated by S. Laufer (University of Tübingen, Germany) as described previously [25]. …”

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Skepinone-L, a Novel Potent and Highly Selective Inhibitor of p38 MAP Kinase, Effectively Impairs Platelet Activation and Thrombus Formation

Borst

Walker²,

Münzer³

et al. 2013

Cell Physiol Biochem

1,339

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“…Where indicated, TGFß1 (60 ng/ml, Sigma, Taufkirchen, Germany), p38 kinase inhibitor Skepinone-L [31] (1 µM, Merck), SGK1 inhibitor GSK-650394 (10 µM, Tocris), or NFκB inhibitor Wogonin (100 µM, Sigma) were added to the medium.…”

Section: Methodsmentioning

confidence: 99%

P38 Kinase, SGK1 and NF-κB Dependent Up-Regulation of Na+/Ca2+ Exchanger Expression and Activity Following TGFß1 Treatment of Megakaryocytes

Al‐Maghout

Pelzl

Sahu³

et al. 2017

Cell Physiol Biochem

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Background: TGFβ1, a decisive regulator of megakaryocyte maturation and platelet formation, has previously been shown to up-regulate both, store operated Ca²⁺ entry (SOCE) and Ca²⁺ extrusion by Na⁺/Ca²⁺ exchange. The growth factor thus augments the increase of cytosolic Ca²⁺ activity ([Ca²⁺]_i) following release of Ca²⁺ from intracellular stores and accelerates the subsequent decline of [Ca²⁺]_i. The effect on SOCE is dependent on a signaling cascade including p38 kinase, serum & glucocorticoid inducible kinase SGK1, and nuclear factor NFκB. The specific Na⁺/Ca²⁺ exchanger isoforms involved and the signalling regulating the Na⁺/Ca²⁺ exchangers remained, however elusive. The present study explored, whether TGFβ1 influences the expression and function of K⁺ insensitive (NCX) and K⁺ sensitive (NCKX) Na⁺/Ca²⁺ exchangers, and aimed to shed light on the signalling involved. Methods: In human megakaryocytic cells (MEG01) RT-PCR was performed to quantify NCX/NCKX isoform transcript levels, [Ca²⁺]_i was determined by Fura-2 fluorescence, and Na⁺/Ca²⁺ exchanger activity was estimated from the increase of [Ca²⁺]_i following switch from an extracellular solution with 130 or 90 mM Na⁺ and 0 mM Ca²⁺ to an extracellular solution with 0 Na⁺ and 2 mM Ca²⁺. K⁺ concentration was 0 mM for analysis of NCX and 40 mM for analysis of NCKX. Results: TGFβ1 (60 ng/ml, 24 h) significantly increased the transcript levels of NCX1, NCKX1, NCKX2 and NCKX5. Moreover, TGFβ1 (60 ng/ml, 24 h) significantly increased the activity of both, NCX and NCKX. The effect of TGFβ1 on NCX and NCKX transcript levels and activity was significantly blunted by p38 kinase inhibitor Skepinone-L (1 µM), the effect on NCX and NCKX activity further by SGK1 inhibitor GSK-650394 (10 µM) and NFκB inhibitor Wogonin (100 µM). Conclusions: TGFβ1 markedly up-regulates transcription of NCX1, NCKX1, NCKX2, and NCKX5 and thus Na⁺/Ca²⁺ exchanger activity, an effect requiring p38 kinase, SGK1 and NFκB.

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Inferential NMR/X‐ray‐Based Structure Determination of a Dibenzo[a,d]cycloheptenone Inhibitor–p38α MAP Kinase Complex in Solution

et al. 2012

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Complex problem: The crystal structure of p38α mitogen‐activated protein kinase in complex with a dibenzo[a,d]cycloheptenone inhibitor was found to be incompatible with NMR data of the same complex in solution. By using inferential structure determination (ISD) with restraints from X‐ray crystallography and NMR spectra, a structure that is compatible with both data sets and very close to the X‐ray crystal structure was generated (see picture).

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Skepinone-L is a selective p38 mitogen-activated protein kinase inhibitor

Cited by 110 publications

References 25 publications

Skepinone-L, a Novel Potent and Highly Selective Inhibitor of p38 MAP Kinase, Effectively Impairs Platelet Activation and Thrombus Formation

Skepinone-L, a Novel Potent and Highly Selective Inhibitor of p38 MAP Kinase, Effectively Impairs Platelet Activation and Thrombus Formation

P38 Kinase, SGK1 and NF-κB Dependent Up-Regulation of Na+/Ca2+ Exchanger Expression and Activity Following TGFß1 Treatment of Megakaryocytes

Inferential NMR/X‐ray‐Based Structure Determination of a Dibenzo[a,d]cycloheptenone Inhibitor–p38α MAP Kinase Complex in Solution

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