Pharmacological evidence for complex and multiple site interaction of CXCR4 with SDF-1alpha: implications for development of selective CXCR4 antagonists

Immunol Lett. 2001 Aug 1;78(1):29-34. doi: 10.1016/s0165-2478(01)00228-0.


The C-X-C chemokine SDF-1 and its receptor CXCR4, mediate a pivotal role in the pathophysiology of HIV-1 infection and vascular inflammatory diseases. In this study, we investigated the pharmacological properties of SDF-1alpha interaction with CXCR4 in human leukemia cell lines. Our data, based on [125I]-SDF-1alpha radioligand binding, SDF-1alpha-induced [35S]-GTPgammaS binding and use of specific CXCR4 antagonist AMD3100 reveals the complex nature of SDF-1alpha-CXCR4 interaction. Firstly, homologous competition with cold SDF-1alpha revealed a bimodal ligand displacement curve and secondly, although AMD3100 inhibited both SDF-1alpha-mediated chemotaxis (IC(50)=4.7 nM) and [35S]-GTPgammaS binding (IC(50)=7.4 nM) with high affinity, it was intriguingly up to 3000-fold less potent (IC(50)=15.2 microM) in the radioligand binding assay. These results provide pharmacological evidence for the recently described two-site model for SDF-1alpha-CXCR4 interaction. Accordingly, inhibition of SDF-1alpha binding to one of the receptor sites is sufficient to antagonize function, without causing its complete displacement from the receptor. Furthermore, these findings have important implications in the development and evaluation of CXCR4-selective small molecule antagonists for therapeutic use.

MeSH terms

  • Benzylamines
  • Binding, Competitive / drug effects
  • Chemokine CXCL12
  • Chemokines, CXC / chemistry
  • Chemokines, CXC / immunology
  • Chemokines, CXC / metabolism*
  • Chemotaxis / drug effects
  • Cyclams
  • Dose-Response Relationship, Immunologic
  • HL-60 Cells
  • Heterocyclic Compounds / pharmacology
  • Humans
  • Jurkat Cells
  • Models, Molecular
  • Protein Binding / drug effects
  • Receptors, CXCR4 / antagonists & inhibitors
  • Receptors, CXCR4 / chemistry
  • Receptors, CXCR4 / metabolism*
  • Receptors, Cell Surface
  • Stromal Cells
  • Structure-Activity Relationship


  • Benzylamines
  • CXCL12 protein, human
  • Chemokine CXCL12
  • Chemokines, CXC
  • Cyclams
  • Heterocyclic Compounds
  • Receptors, CXCR4
  • Receptors, Cell Surface
  • plerixafor