Antagonists of GPR35 display high species ortholog selectivity and varying modes of action

Jenkins, L. , Harries, N., Lappin, J.E., MacKenzie, A.E., Neetoo-Isseljee, Z., Southern, C., McIver, E.G., Nicklin, S.A. , Taylor, D.L. and Milligan, G. (2013) Antagonists of GPR35 display high species ortholog selectivity and varying modes of action. Journal of Pharmacology and Experimental Therapeutics, 343(3), pp. 683-695. (doi:10.1124/jpet.112.198945)

Jenkins, L. , Harries, N., Lappin, J.E., MacKenzie, A.E., Neetoo-Isseljee, Z., Southern, C., McIver, E.G., Nicklin, S.A. , Taylor, D.L. and Milligan, G. (2013) Antagonists of GPR35 display high species ortholog selectivity and varying modes of action. Journal of Pharmacology and Experimental Therapeutics, 343(3), pp. 683-695. (doi:10.1124/jpet.112.198945)

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Publisher's URL: http://dx.doi.org/10.1124/jpet.112.198945

Abstract

Variation in pharmacology and function of ligands at species orthologs can be a confounding feature in understanding the biology and role of poorly characterized receptors. Substantial selectivity in potency of a number of GPR35 agonists has previously been demonstrated between human and rat orthologs of this G protein-coupled receptor. Via a bioluminescence resonance energy transfer-based assay of induced interactions between GPR35 and β-arrestin-2, addition of the mouse ortholog to such studies indicated that, as for the rat ortholog, murine GPR35 displayed very low potency for pamoate, whereas potency for the reference GPR35 agonist zaprinast was intermediate between the rat and human orthologs. This pattern was replicated in receptor internalization and G protein activation assays. The effectiveness and mode of action of two recently reported GPR35 antagonists, methyl-5-[(tert-butylcarbamothioylhydrazinylidene)methyl]-1-(2,4-difluorophenyl)pyrazole-4-carboxylate (CID-2745687) and 2-hydroxy-4-[4-(5Z)-5-[(E)-2-methyl-3-phenylprop-2-enylidene]-4-oxo-2-sulfanylidene-1,3-thiazolidin-3-yl]butanoylamino)benzoic acid (ML-145), were investigated. Both CID-2745687 and ML-145 competitively inhibited the effects at human GPR35 of cromolyn disodium and zaprinast, two agonists that share an overlapping binding site. By contrast, although ML-145 also competitively antagonized the effects of pamoate, CID-2745687 acted in a noncompetitive fashion. Neither ML-145 nor CID-2745687 was able to effectively antagonize the agonist effects of either zaprinast or cromolyn disodium at either rodent ortholog of GPR35. These studies demonstrate that marked species selectivity of ligands at GPR35 is not restricted to agonists and considerable care is required to select appropriate ligands to explore the function of GPR35 in nonhuman cells and tissues.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Harries, Mr Nicholas and Milligan, Professor Graeme and Nicklin, Professor Stuart and Jenkins, Mrs Laura
Authors: Jenkins, L., Harries, N., Lappin, J.E., MacKenzie, A.E., Neetoo-Isseljee, Z., Southern, C., McIver, E.G., Nicklin, S.A., Taylor, D.L., and Milligan, G.
College/School:College of Medical Veterinary and Life Sciences > Institute of Cardiovascular and Medical Sciences
College of Medical Veterinary and Life Sciences > Institute of Molecular Cell and Systems Biology
Journal Name:Journal of Pharmacology and Experimental Therapeutics
Journal Abbr.:JPET
Publisher:American Society for Pharmacology and Experimental Therapeutics
ISSN:0022-3565
ISSN (Online):1521-0103
Published Online:11 September 2012

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
500152Doctoral Training Grant 2009-15Timothy PalmerBiotechnology and Biological Sciences Research Council (BBSRC)BB/F016735/1RI CARDIOVASCULAR & MEDICAL SCIENCES
492451BHF 4 Year PhD ProgrammeAnna DominiczakBritish Heart Foundation (BHF)FS/09/052RI CARDIOVASCULAR & MEDICAL SCIENCES