Receptor binding mode and pharmacological characterization of a potent and selective dual CXCR1/CXCR2 non-competitive allosteric inhibitor.

Abstract

BACKGROUND AND PURPOSE:

DF 2156A is a new dual inhibitor of IL-8 receptors CXCR1 and CXCR2 with an optimal pharmacokinetic profile. We characterized its binding mode, molecular mechanism of action and selectivity, and evaluated its therapeutic potential.

EXPERIMENTAL APPROACH:

The binding mode, molecular mechanism of action and selectivity were investigated using chemotaxis of L1.2 transfectants and human leucocytes, in addition to radioligand and [(35) S]-GTPγS binding approaches. The therapeutic potential of DF 2156A was evaluated in acute (liver ischaemia and reperfusion) and chronic (sponge-induced angiogenesis) experimental models of inflammation.

KEY RESULTS:

A network of polar interactions stabilized by a direct ionic bond between DF 2156A and Lys(99) on CXCR1 and the non-conserved residue Asp(293) on CXCR2 are the key determinants of DF 2156A binding. DF 2156A acted as a non-competitive allosteric inhibitor blocking the signal transduction leading to chemotaxis without altering the binding affinity of natural ligands. DF 2156A effectively and selectively inhibited CXCR1/CXCR2-mediated chemotaxis of L1.2 transfectants and leucocytes. In a murine model of sponge-induced angiogenesis, DF 2156A reduced leucocyte influx, TNF-α production and neovessel formation. In vitro, DF 2156A prevented proliferation, migration and capillary-like organization of HUVECs in response to human IL-8. In a rat model of liver ischaemia and reperfusion (I/R) injury, DF 2156A decreased PMN and monocyte-macrophage infiltration and associated hepatocellular injury.

CONCLUSION AND IMPLICATIONS:

DF 2156A is a non-competitive allosteric inhibitor of both IL-8 receptors CXCR1 and CXCR2. It prevented experimental angiogenesis and hepatic I/R injury in vivo and, therefore, has therapeutic potential for acute and chronic inflammatory diseases.

• Authors: R Bertini, L S Barcelos, A R Beccari, B Cavalieri, A Moriconi, C Bizzarri, P Di Benedetto, C Di Giacinto, I Gloaguen, E Galliera, M M Corsi, R C Russo, S P Andrade, M C Cesta, G Nano, A Aramini, J C Cutrin, M Locati, M Allegretti, M M Teixeira
• Journal: British journal of pharmacology (Br J Pharmacol) Vol. 165 Issue 2 Pg. 436-54 (Jan 2012) ISSN: 1476-5381 [Electronic] England
• PMID: 21718305 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2011 The Authors. British Journal of Pharmacology © 2011 The British Pharmacological Society.

Chemical References

• 2'-((4'-trifluoromethanesulfonyloxy)phenyl)-N-methanesulfonylpropionamide
• Anti-Inflammatory Agents
• Interleukin-8
• Receptors, Interleukin-8A
• Receptors, Interleukin-8B
• Sulfonamides

Topics

• Animals
• Anti-Inflammatory Agents (pharmacokinetics, pharmacology, therapeutic use)
• Cell Membrane (metabolism)
• Cell Proliferation (drug effects)
• Chemotaxis, Leukocyte (drug effects)
• Disease Models, Animal
• Human Umbilical Vein Endothelial Cells (drug effects, metabolism)
• Humans
• Interleukin-8 (metabolism)
• Leukocytes (drug effects, immunology, metabolism)
• Liver (drug effects, immunology, pathology)
• Male
• Mice
• Mice, Inbred C57BL
• Models, Molecular
• Mutagenesis, Site-Directed
• Neovascularization, Pathologic (drug therapy, metabolism, pathology)
• Rats
• Rats, Sprague-Dawley
• Receptors, Interleukin-8A (antagonists & inhibitors, genetics, metabolism)
• Receptors, Interleukin-8B (antagonists & inhibitors, genetics, metabolism)
• Reperfusion Injury (drug therapy, immunology, pathology)
• Skin (blood supply)
• Sulfonamides (pharmacokinetics, pharmacology, therapeutic use)