TY - JOUR T1 - Biochemical, Cellular, and Anti-Inflammatory Properties of a Potent, Selective, Orally Bioavailable Benzamide Inhibitor of Rho Kinase Activity JF - Journal of Pharmacology and Experimental Therapeutics JO - J Pharmacol Exp Ther SP - 707 LP - 716 DO - 10.1124/jpet.110.166033 VL - 333 IS - 3 AU - Lakshman E. Rajagopalan AU - Michael S. Davies AU - Larry E. Kahn AU - Christine M. Kornmeier AU - Hideaki Shimada AU - Toni A. Steiner AU - Ben S. Zweifel AU - Jay M. Wendling AU - Maria A. Payne AU - Richard F. Loeffler AU - Brenda L. Case AU - Monica B. Norton AU - Mihir D. Parikh AU - Olga V. Nemirovskiy AU - Robert J. Mourey AU - Jaime L. Masferrer AU - Thomas P. Misko AU - Stephen A. Kolodziej Y1 - 2010/06/01 UR - http://jpet.aspetjournals.org/content/333/3/707.abstract N2 - Rho kinase, is the most widely studied downstream effector of the small Rho GTPase RhoA. Two Rho kinase isoforms have been described and are frequently referred to in the literature as ROCK1and ROCK2. The RhoA–Rho kinase pathway has been implicated in the recruitment of cellular infiltrates to disease loci in a number of preclinical animal models of inflammatory disease. In this study, we used biochemical enzyme assays and a cellular target biomarker assay to define PF-4950834 [N-methyl-3-{[(4-pyridin-4-ylbenzoyl)amino]methyl}benzamide] as an ATP-competitive, selective Rho kinase inhibitor. We further used PF-4950834 to study the role of Rho kinase activation in lymphocyte and neutrophil migration in addition to the endothelial cell-mediated expression of adhesion molecules and chemokines, which are essential for leukocyte recruitment. The inhibitor blocked stromal cell-derived factor-1α-mediated chemotaxis of T lymphocytes in vitro and the synthesis of vascular cell adhesion molecule-1 and intercellular adhesion molecule-1 in activated human endothelial cells in vitro. The secretion of chemokines interleukin-8 and monocyte chemoattractant protein-1 was also inhibited in activated endothelial cells. In addition, when dosed orally, the compound potently inhibited neutrophil migration in a carrageenan-induced acute inflammation model. In summary, we have used a pharmacologic approach to link Rho kinase activation to multiple phenotypes that can contribute to leukocyte infiltration. Inhibition of this pathway therefore could be strongly anti-inflammatory and provide therapeutic benefit in chronic inflammatory diseases. Copyright © 2010 by The American Society for Pharmacology and Experimental Therapeutics ER -