A Benzothiophene Inhibitor of MAPK-Activated Protein Kinase 2 (MK2) Inhibits TNFα Production and has Oral Anti-Inflammatory Efficacy in Acute and Chronic Models of Inflammation.

Activation of the p38 kinase pathway in immune cells leads to the transcriptional and translational regulation of pro-inflammatory cytokines. MAPKAP kinase 2 (MK2), a direct downstream substrate of p38 kinase, regulates LPS-stimulated TNFα and IL-6 production through modulating the stability and translation of these mRNAs. Developing small molecule inhibitors of MK2 may yield anti-inflammatory efficacy with a different safety profile relative to p38 kinase inhibitors. This report describes the pharmacologic properties of a benzothiophene MK2 inhibitor [(10R)-10-methyl-3-(6-methylpyridin-3-yl)-9,10,11,12-tetrahydro-8H-[1,4]diazepino[59,69:4,5]thieno[3,2-f]quinolin-8-one; PF-3644022]. PF-3644022 is a potent freely reversible ATP-competitive compound that inhibits MK2 kinase activity (K i = 3 nM) with good selectivity when profiled against 200 human kinases. In the human U937 monocytic cell line or PBMCs, PF-3644022 potently inhibits TNFα production with similar activity (IC 50 = 160 nM). PF-3644022 blocks TNFα and IL-6 production in LPS-stimulated human whole blood with IC 50 values of 1.6 μM and 10.3 μM, respectively. Inhibition of TNFα in U937 cells and blood correlates closely with inhibition of phospho-HSP27, a target biomarker of MK2 activity. PF-3644022 displays good pharmacokinetic parameters in rats and is orally efficacious in both the rat acute LPS-induced TNFα model and the chronic streptococcal cell wall-induced arthritis model. Dose-dependent inhibition of TNFα production in the acute model, and inhibition of paw swelling in the chronic model is observed with ED 50 values of 6.9 and 20 mg/kg, respectively. PF-3644022 efficacy in the chronic inflammation model is strongly correlated with maintaining a C min greater than the EC 50 measured in the rat LPS-induced TNFα model.