title: Antagonism of human CC-chemokine receptor 4 can be achieved through three distinct binding sites on the receptor.
abstract: Chemokine receptor antagonists appear to access two distinct binding sites on different members of this receptor family. One class of CCR4 antagonists has been suggested to bind to a site accessible from the cytoplasm while a second class did not bind to this site. In this report, we demonstrate that antagonists representing a variety of structural classes bind to two distinct allosteric sites on CCR4. The effects of pairs of low-molecular weight and/or chemokine CCR4 antagonists were evaluated on CCL17- and CCL22-induced responses of human CCR4(+) T cells. This provided an initial grouping of the antagonists into sets which appeared to bind to distinct binding sites. Binding studies were then performed with radioligands from each set to confirm these groupings. Some novel receptor theory was developed to allow the interpretation of the effects of the antagonist combinations. The theory indicates that, generally, the concentration-ratio of a pair of competing allosteric modulators is maximally the sum of their individual effects while that of two modulators acting at different sites is likely to be greater than their sum. The low-molecular weight antagonists could be grouped into two sets on the basis of the functional and binding experiments. The antagonistic chemokines formed a third set whose behaviour was consistent with that of simple competitive antagonists. These studies indicate that there are two allosteric regulatory sites on CCR4.
authors.affiliation: Lead Optimisation, Respiratory CEDD, GlaxoSmithKline Gunnels Wood Road, Stevenage, Herts, SG1 2NY, UK.
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Slack
authors.affiliation: Lead Optimisation, Respiratory CEDD, GlaxoSmithKline Gunnels Wood Road, Stevenage, Herts, SG1 2NY, UK.
authors.forename: Linda J
authors.initials: LJ
authors.lastname: Russell
authors.affiliation: Computational Chemistry, GlaxoSmithKline Gunnels Wood Road, Stevenage, Herts, SG1 2NY, UK.
authors.forename: Nick P
authors.initials: NP
authors.lastname: Barton
authors.affiliation: Lead Optimisation, Respiratory CEDD, GlaxoSmithKline Gunnels Wood Road, Stevenage, Herts, SG1 2NY, UK.
authors.forename: Cathryn
authors.initials: C
authors.lastname: Weston
authors.affiliation: Lead Optimisation, Respiratory CEDD, GlaxoSmithKline Gunnels Wood Road, Stevenage, Herts, SG1 2NY, UK.
authors.forename: Giovanna
authors.initials: G
authors.lastname: Nalesso
authors.affiliation: Lead Optimisation, Respiratory CEDD, GlaxoSmithKline Gunnels Wood Road, Stevenage, Herts, SG1 2NY, UK.
authors.forename: Sally-Anne
authors.initials: SA
authors.lastname: Thompson
authors.affiliation: Biological Reagents and Assay Development, GlaxoSmithKline Gunnels Wood Road, Stevenage, Herts, SG1 2NY, UK.
authors.forename: Morven
authors.initials: M
authors.lastname: Allen
authors.affiliation: Biological Reagents and Assay Development, GlaxoSmithKline Gunnels Wood Road, Stevenage, Herts, SG1 2NY, UK.
authors.forename: Yu Hua
authors.initials: YH
authors.lastname: Chen
authors.affiliation: Biological Reagents and Assay Development, GlaxoSmithKline Gunnels Wood Road, Stevenage, Herts, SG1 2NY, UK.
authors.forename: Ashley
authors.initials: A
authors.lastname: Barnes
authors.affiliation: Medicinal Chemistry, Respiratory CEDD, GlaxoSmithKline Gunnels Wood Road, Stevenage, Herts, SG1 2NY, UK.
authors.forename: Simon T
authors.initials: ST
authors.lastname: Hodgson
authors.affiliation: Lead Optimisation, Respiratory CEDD, GlaxoSmithKline Gunnels Wood Road, Stevenage, Herts, SG1 2NY, UK.
authors.forename: David A
authors.initials: DA
authors.lastname: Hall
keywords: Allosteric modulator; CCL17; CCL22; CCR4; CCR4 antagonist; T cell; chemokine