Improved lymphocyte function-associated antigen-1 (LFA-1) inhibition by statin derivatives: molecular basis determined by x-ray analysis and monitoring of LFA-1 conformational changes in vitro and ex vivo

J Biol Chem. 2004 Nov 5;279(45):46764-71. doi: 10.1074/jbc.M407951200. Epub 2004 Aug 10.

Abstract

The integrin lymphocyte function-associated antigen-1 (LFA-1) (alphaLbeta2; CD11a/CD18) plays an important role in leukocyte migration and T cell activation. LFA-1 is inhibited by the cholesterol-lowering drug lovastatin, which binds to an allosteric site of the alphaL I domain termed the lovastatin site (L-site). Here we report for the first time the x-ray structures of the LFA-1 I domain complexed with derivatives of lovastatin optimized for LFA-1 inhibition. This analysis identified two new subpockets within the L-site occupied by chemical groups of the statin derivatives but not by lovastatin itself. Occupancy of these L-site subpockets led to distinct conformational changes in LFA-1, which were detectable by an epitope-monitoring assay. We utilized this assay to demonstrate improved LFA-1 inhibition in human blood in vitro and in blood samples from treated animals ex vivo. Moreover, we demonstrate that the novel lovastatin-derived LFA-1 inhibitor LFA878 exhibits potent anti-inflammatory effects in carrageenan-induced rat paw edema. In summary, the findings reported here extend the understanding of LFA-1 inhibition at the molecular level, allow for the identification and design of LFA-1 inhibitors of further enhanced potency, and support the expectation that LFA-1 inhibitors binding to the L-site will be of therapeutic value in treating inflammatory diseases.

MeSH terms

  • Animals
  • Binding Sites
  • Blood / metabolism
  • Carrageenan / pharmacology
  • Cell Movement
  • Dose-Response Relationship, Drug
  • Edema
  • Epitopes / chemistry
  • Female
  • Humans
  • Hydroxymethylglutaryl-CoA Reductase Inhibitors / pharmacology*
  • Inhibitory Concentration 50
  • Jurkat Cells
  • Leukocytes / cytology
  • Lovastatin / pharmacology
  • Lymphocyte Function-Associated Antigen-1 / chemistry*
  • Lymphocyte Function-Associated Antigen-1 / metabolism
  • Male
  • Models, Chemical
  • Models, Molecular
  • Naphthalenes / chemistry*
  • Naphthalenes / pharmacology*
  • Oxazines / chemistry*
  • Oxazines / pharmacology*
  • Protein Binding
  • Protein Conformation
  • Protein Structure, Tertiary
  • Rabbits
  • Rats
  • T-Lymphocytes / cytology
  • Time Factors
  • X-Rays

Substances

  • Epitopes
  • Hydroxymethylglutaryl-CoA Reductase Inhibitors
  • LFA 878
  • Lymphocyte Function-Associated Antigen-1
  • Naphthalenes
  • Oxazines
  • Carrageenan
  • Lovastatin

Associated data

  • PDB/1XDD
  • PDB/1XDG