title: Characterization of the molecular pharmacology of AMD3100: a specific antagonist of the G-protein coupled chemokine receptor, CXCR4.
abstract: The chemokine receptor CXCR4 is widely expressed on different cell types, is involved in leukocyte chemotaxis, and is a co-receptor for HIV. AMD3100 has been shown to be a CXCR4 receptor antagonist, and to block HIV infection of T-tropic, X4-using, virus in vitro and in vivo. AMD3100 is an effective mobilizer of hematopoietic stem cells and is being investigated in clinical trials in multiple myeloma and non-Hodgkins lymphoma patients. Using the CCRF-CEM T-cell line that constitutively expresses CXCR4 we confirmed that AMD3100 was an antagonist of SDF-1/CXCL12 ligand binding (IC50=651+/-37 nM). We have also shown that AMD3100 inhibits SDF-1 mediated GTP-binding (IC50=27+/-2.2 nM), SDF-1 mediated calcium flux (IC50=572+/-190 nM), and SDF-1 stimulated chemotaxis (IC50=51+/-17 nM). AMD3100 did not inhibit calcium flux against cells expressing CXCR3, CCR1, CCR2b, CCR4, CCR5 or CCR7 when stimulated with their cognate ligands, nor did it inhibit receptor binding of LTB4. AMD3100 did not, on its own, induce a calcium flux in the CCRF-CEM cells, which express multiple GPCRs including CXCR4, CCR4 and CCR7. Furthermore, AMD3100 neither stimulated GTP-binding, an assay for GPCR activation, in CEM cell membranes; nor chemotaxis of CCRF-CEM cells. These data therefore demonstrate that AMD3100 is a specific antagonist of CXCR4, is not cross-reactive with other chemokine receptors, and is not an agonist of CXCR4.
authors.affiliation: AnorMED Inc., #200 20353 64th Avenue, Langley, BC V2Y 1N5, Canada. sfricker@anormed.com
authors.forename: Simon P
authors.initials: SP
authors.lastname: Fricker
authors.forename: Virginia
authors.initials: V
authors.lastname: Anastassov
authors.forename: Jennifer
authors.initials: J
authors.lastname: Cox
authors.forename: Marilyn C
authors.initials: MC
authors.lastname: Darkes
authors.forename: Ognjen
authors.initials: O
authors.lastname: Grujic
authors.forename: Stefan R
authors.initials: SR
authors.lastname: Idzan
authors.forename: Jean
authors.initials: J
authors.lastname: Labrecque
authors.forename: Gloria
authors.initials: G
authors.lastname: Lau
authors.forename: Renee M
authors.initials: RM
authors.lastname: Mosi
authors.forename: Kim L
authors.initials: KL
authors.lastname: Nelson
authors.forename: Ling
authors.initials: L
authors.lastname: Qin
authors.forename: Zeffy
authors.initials: Z
authors.lastname: Santucci
authors.forename: Rebecca S Y
authors.initials: RS
authors.lastname: Wong
mesh_terms: D001596:Benzylamines; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D054377:Chemokine CXCL12; D019743:Chemokines, CXC / Q000378:metabolism; D002633:Chemotaxis / Q000187:drug effects; D000080027:Cyclams; D006571:Heterocyclic Compounds / Q000494:pharmacology*; D006801:Humans; D011485:Protein Binding; D019718:Receptors, CXCR4 / Q000037:antagonists & inhibitors*; D015398:Signal Transduction / Q000187:drug effects
chemical_list: D001596:Benzylamines; C515255:CXCL12 protein, human; D054377:Chemokine CXCL12; D019743:Chemokines, CXC; D000080027:Cyclams; D006571:Heterocyclic Compounds; D019718:Receptors, CXCR4; C088327:plerixafor; D002118:Calcium