Identification of a selective thieno[2,3-c]pyridine inhibitor of COT kinase and TNF-α production

Cot/tpl2 (also known as MAP3K8) has emerged as a new and potentially interesting therapeutic anti-inflammatory target. Here, we report the first study of Cot/tpl2 involvement in acute peripheral inflammation in vivo. Six hours after an intraplantar injection of zymosan, Cot/tpl2 ؊/؊ mice showed a 47% reduction in myeloperoxidase activity, concomitant with a 46% lower neutrophil recruitment and a 40% decreased luminol-mediated bioluminescence imaging in vivo. Accordingly, Cot/tpl2 deficiency provoked a 25-30% reduction in luminolmediated bioluminescence and neutrophil recruitment together with a 65% lower macrophage recruitment 4 h following zymosan-induced peritonitis. Significantly impaired levels of G-CSF and GM-CSF and of other cytokines such as TNF␣, IL-1␤, and IL-6, as well as some chemokines such as MCP-1, MIP-1␤, and keratinocyte-derived chemokine, were detected during the acute zymosan-induced intraplantar inflammatory response in Cot/ tpl2 ؊/؊ mice. Moreover, Cot/tpl2 deficiency dramatically decreased the production of the hypernociceptive ligand NGF at the inflammatory site during the course of inflammation. Most importantly, Cot/tpl2 deficiency significantly reduced zymosan-induced inflammatory hypernociception in mice, with a most pronounced effect of a 50% decrease compared with wild type (WT) at 24 h following intraplantar injection of zymosan. At this time, Cot/ tpl2 ؊/؊ mice showed significantly reduced NGF, TNF␣, and prostaglandin E 2 levels compared with WT littermates. In conclusion, our study demonstrates an important role of Cot/tpl2 in the NGF, G-CSF, and GM-CSF production and myeloperoxidase activity in the acute inflammatory response process and its implication in inflammatory hypernociception. 2). Cot/tpl2, also known as MAP3K8, is the sole MAP3K that activates the MKK1/ 2-ERK1/2 pathway in response to stimulation of the TLR/IL-1 receptor superfamily, as well as in response to the activation of some receptors of the TNF family (3-10). However, Cot/tpl2 does not participate in the activation of ERK1/2 by stimulation of the C-type lectin receptor dectin-1 (11). In resting cells, Cot/tpl-2 forms a stable and inactive complex with p105 NF-B and ABIN2 (A20-binding inhibitor of NF-B2), among other proteins, to protect Cot/tpl-2 from degradation. Adequate TLR/IL-1 receptor stimulation induces the activation of the IKK complex; active IKK␤ kinase phosphorylates p105 NF-B, triggering its partial degradation to p50 NF-B (12-15). Cot/tpl2 is then dissociated from the complex and with an adequate phosphorylation state (16 -19) is capable of activating MKK1 and consequently ERK1/2 (6, 20 -22) prior to being rapidly degraded through the proteasome pathway (6,20,23). Cot/tpl2 is required to process pre-TNF␣ to its mature secreted form in LPS-stimulated macrophages (24). In addition, in different isolated cell types Cot/tpl2 controls the secretion of other cytokines and chemokines such as IL-8, MCP-1, MIP-1␤, KC,10,[24][25][26][27]. Moreover, Cot/tpl2 activation is necessary for the production of PGE 2 ...

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confidence: 99%

Cot/tpl2 (MAP3K8) Mediates Myeloperoxidase Activity and Hypernociception following Peripheral Inflammation*

Soria-Castro

Krzyzanowska

Pelaéz

et al. 2010

“…Furthermore, we and others have demonstrated that Tpl2 is important in transducing signals downstream of multiple TLRs, leading to ERK activation and the expression of a subset of proinflammatory mediators, including TNF (24 -26). Consequently, Tpl2 small molecule inhibitors are now being developed as a potential treatment for chronic autoimmune conditions, such as rheumatoid arthritis, in which TNF plays a pathologic role (27)(28)(29)(30).…”

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confidence: 99%

Tumor Progression Locus 2 (Tpl2) Kinase Promotes Chemokine Receptor Expression and Macrophage Migration during Acute Inflammation

Rowley

Kuriakose

Dockery

et al. 2014

Background:The accumulation of activated leukocytes correlates with autoimmunity and is regulated by chemokines. Results: Tpl2-deficient macrophages display impaired chemokine receptor expression and migration under inflammatory conditions. Conclusion: Tpl2 promotes the induction and maintenance of macrophage chemokine receptor expression and cellular migration in vivo. Significance: Tpl2 inhibition may represent a treatment for autoimmune disorders by modulating chemokine receptor expression and preventing local accumulation of activated macrophages.

“…The discovery of small molecule inhibitors in high throughput screens using COT kinase has been described (15)(16)(17)(18)(19)(20)(21)(22). Although some progress in the advancement of these molecules has been made, the low sequence homology of COT kinase to other serine-threonine (ST) kinases and the lack of structural information have hampered rapid progress in the understanding of the structure-activity relationship of the reported compounds.…”

Section: Discussionmentioning

confidence: 99%

“…Conservation of between groups of strongly similar and weakly similar properties are indicated with a colon or a period, respectively. tivity relationship of COT kinase inhibitors (15)(16)(17)(18)(19)(20)(21)(22)40) and reduces the predictability of structure-guided medicinal chemistry approaches, such as scaffold morphing and fragment growing. Although the presented co-crystal structures of COT kinase inhibitor complexes provided a first glimpse on the binding mode of early hits, the high throughput in vitro phosphorylation assay we report above became an invaluable tool to rapidly test newly synthesized derivatives for potency and selectivity.…”

Section: Discussionmentioning

confidence: 99%

“…Although the discovery of COT inhibitors has been reported in the literature (15)(16)(17)(18)(19)(20)(21)(22), difficulties in obtaining monodisperse protein and the lack of structural information have hampered the discovery of chemical compounds suitable for clinical studies. Careful selection of protein constructs and optimizations in the protein production procedures allowed us to obtain pure enzyme of exquisite biochemical activity.…”

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confidence: 99%

See 1 more Smart Citation

The Crystal Structure of Cancer Osaka Thyroid Kinase Reveals an Unexpected Kinase Domain Fold

Gutmann

Hinniger

Fendrich

et al. 2015

Background: Cancer Osaka thyroid (COT) kinase plays a crucial role in inflammatory diseases and cancer. Results: Production of catalytically competent COT kinase yielded protein suitable for structure guided drug discovery. Conclusion: COT kinase has a unique and structurally versatile active site. Significance: The discovery of a novel variation of the protein kinase fold will impact drug discovery for COT kinase.