Structure Function Differences in Nonpeptide CCR1 Antagonists for Human and Mouse CCR1

Onuffer, James; McCarrick, Margaret A.; Dunning, Laura; Liang, Meina; Rosser, Mary P.; Wei, Guo-Ping; Ng, Howard P.; Horuk, Richard

doi:10.4049/jimmunol.170.4.1910

Cited by 27 publications

(14 citation statements)

References 20 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…2). These mutants included key residues predicted to be in the binding pocket for BX 471 (Tyr-113, Tyr-114, and Ile-259) together with residues Tyr-41, Glu-287, and Tyr-291, identified as playing a role in antagonist binding in previous studies with charged CCR1 antagonists (18,26). Cells separately transfected with either wild-type CCR1 or with point mutants of CCR1 predicted to be outside the antagonist binding site were included as controls (Fig.…”

Section: Resultsmentioning

confidence: 99%

Predictions of CCR1 Chemokine Receptor Structure and BX 471 Antagonist Binding Followed by Experimental Validation

Vaidehi

Schlyer²,

Trabanino

et al. 2006

Journal of Biological Chemistry

Self Cite

114

View full text Add to dashboard Cite

A major challenge in the application of structure-based drug design methods to proteins belonging to the superfamily of G protein-coupled receptors (GPCRs) is the paucity of structural information (1). The 19 chemokine receptors, belonging to the Class A family of GPCRs, are important drug targets not only for autoimmune diseases like multiple sclerosis but also for the blockade of human immunodeficiency virus type 1 entry (2). Using the MembStruk computational method (3), we predicted the three-dimensional structure of the human CCR1 receptor. In addition, we predicted the binding site of the small molecule CCR1 antagonist BX 471, which is currently in Phase II clinical trials (4). Based on the predicted antagonist binding site we designed 17 point mutants of CCR1 to validate the predictions. Subsequent competitive ligand binding and chemotaxis experiments with these mutants gave an excellent correlation to these predictions. In particular, we find that Tyr-113 and Tyr-114 on transmembrane domain 3 and Ile-259 on transmembrane 6 contribute significantly to the binding of BX 471. Finally, we used the predicted and validated structure of CCR1 in a virtual screening validation of the Maybridge data base, seeded with selective CCR1 antagonists. The screen identified 63% of CCR1 antagonists in the top 5% of the hits. Our results indicate that rational drug design for GPCR targets is a feasible approach.Chemokines belong to a large family of small, chemotactic cytokines that regulate the trafficking of immune cells (5) by binding to cell surface receptors belonging to the GPCR 3 superfamily (5). CCR1, the first CC chemokine receptor to be identified, responds to a number of ligands, including MIP-1␣ (CCL3) and RANTES (regulated on activation normal T cell expressed and secreted) (CCL5) (6, 7). The strong association with a wide variety of autoimmune and pro-inflammatory diseases has made the CCR1 protein an attractive therapeutic target, and Berlex has developed a potent, specific, orally available antagonist, BX 471, currently in a Phase II clinical trial (8).The CCR1 antagonist program that yielded the clinical compound BX 471 followed a traditional drug discovery approach starting with high throughput screening of large compound libraries (9). Although high throughput screening is a main pillar of drug-finding programs in the pharmaceutical industry, it has recently been supplemented by in silico methods to maximize the probability of finding attractive novel leads. Structurebased in silico approaches have been challenging for GPCRs, because only one experimental GPCR structure, that of bovine rhodopsin, with only ϳ20% sequence identity to CCR1 (10), has been reported. Recent developments in GPCR structure prediction methods show great potential for structure-based drug design and identifying novel hits from virtual screens (11)(12)(13)(14)(15).In this communication we report a significant test of the computational method MembStruk by predicting the structure of CCR1. Further, we scanned the entire predicted struc...

show abstract

Section: Resultsmentioning

confidence: 99%

Predictions of CCR1 Chemokine Receptor Structure and BX 471 Antagonist Binding Followed by Experimental Validation

Vaidehi

Schlyer²,

Trabanino

et al. 2006

Journal of Biological Chemistry

Self Cite

114

View full text Add to dashboard Cite

show abstract

“…9D). Modeling of the interaction of CCR1 with the hydroxypiperidine BX510 also implicates Tyr-113 and Glu-287 as forming part of the antagonist binding site (39), although in that study there is no role for the Tyr-41 of CCR1 in docking with the compound.…”

Section: Mapping the Site Of Interaction For Ccr1-ucb 35625-mentioning

confidence: 94%

Site-directed Mutagenesis of CC Chemokine Receptor 1 Reveals the Mechanism of Action of UCB 35625, a Small Molecule Chemokine Receptor Antagonist

Mendonça

Fonseca

Phillips

et al. 2005

Journal of Biological Chemistry

View full text Add to dashboard Cite

The chemokine receptor CCR1 and its principal ligand, CCL3/MIP-1␣, have been implicated in the pathology of several inflammatory diseases including rheumatoid arthritis, multiple sclerosis, and asthma. As such, these molecules are the focus of much research with the ultimate aim of developing novel therapies. We have described previously a non-competitive small molecule antagonist of CCR1 (UCB 35625), which we hypothesized interacted with amino acids located within the receptor transmembrane (TM) helices (Sabroe, I., Peck, M. J., Jan Van Keulen, B., Jorritsma, A., Simmons, G., Clapham, P. R., Williams, T. J., and Pease, J. E. (2000) J. Biol. Chem. 275, 25985-25992). Here we describe an approach to identifying the mechanism by which the molecule antagonizes CCR1. Thirty-three point mutants of CCR1 were expressed transiently in L1.2 cells, and the cells were assessed for their capacity to migrate in response to CCL3 in the presence or absence of UCB 35625. Cells expressing the mutant constructs Y41A (TM helix 1, or TM1), Y113A (TM3), and E287A (TM7) were responsive to CCL3 but resistant to the antagonist, consistent with a role for the TM helices in CCR1 interactions with UCB 35625. Subsequent molecular modeling successfully docked the compound with CCR1 and suggests that the antagonist ligates TM1, 2, and 7 of CCR1 and severely impedes access to TM2 and TM3, a region thought to be perturbed by the chemokine amino terminus during the process of receptor activation. Insights into the mechanism of action of these compounds may facilitate the development of more potent antagonists that show promise as future therapeutic agents in the treatment of inflammatory disease.

show abstract

“…Nonpeptide, small molecular, chemokine receptor antagonists sometimes show species specificity (Onuffer et al, 2003). Disparity in the potency of CCR3 antagonism between primates and rodents was more than 60 times based on IC 50 values.…”

Section: Discussionmentioning

confidence: 99%

A Novel, Selective, and Orally Available Antagonist for CC Chemokine Receptor 3

Morokata¹,

Suzuki²,

Masunaga³

et al. 2005

J Pharmacol Exp Ther

View full text Add to dashboard Cite

CC chemokine ligand 11 (CCL11/eotaxin) and other CC chemokine receptor 3 (CCR3) ligands (CCL24/eotaxin-2, CCL26/ eotaxin-3, CCL13/monocyte chemotactic protein-4, etc.) play important roles in the chemotaxis and activation of eosinophils and other CCR3-expressing cells (basophils, mast cells, and CD4 ϩ T helper 2 cells) in allergic inflammation incidents, including asthma and rhinitis. A newly synthesized compound, (1 mg/kg) inhibited CCL11-induced shape change of whole blood eosinophils in cynomolgus monkeys. Intravenous injection of YM-355179 (1 mg/kg) also inhibited eosinophil infiltration into airways of cynomolgus monkeys after segmental bronchoprovocation with CCL11. These results indicate that YM-355179 is a novel, selective, and orally available CCR3 antagonist with therapeutic potential for treating eosinophil-related allergic inflammatory diseases.Eosinophils play a crucial role in allergic diseases such as bronchial asthma and allergic rhinitis (Durham and Kay, 1985;Terada et al., 1994). CCR3, a seven-transmembranespanning G protein-coupled receptor, is a major chemokine receptor expressed on allergic inflammatory cells including not only eosinophils but also basophils, mast cells, and T helper 2-type CD4 ϩ cells (Combadiere et al., 1995;Post et al., 1995;Sabroe et al., 1998). This receptor has been identified in humans, monkeys, mice, rats, and guinea pigs (GarciaZepeda et al., 1996;Forssmann et al., 1997;Sallusto et al., 1997;Zhang et al., 2002). Human CCR3 binds multiple chemokine ligands such as CCL11/eotaxin, CCL24/eotaxin-2, CCL26/eotaxin-3, and CCL13/MCP-4 with high affinity, whereas CCR3 binds CCL5/regulated on activation normal T cell expressed and secreted and CCL7/MCP-3 with lower affinity (Forssmann et al., 1997;Stellato et al., 1997;Doucet et al., 1998). CCR3 ligands are produced by multiple cell types, including lung and mucosal fibroblast cells, bronchial epithelial cells, vascular endothelial cells, smooth muscle cells, macrophages, mast cells, and eosinophils (Lamkhioued et al., 1997;Lilly et al., 1997;Shinkai et al., 1999;Hirst et al., 2002;Menzies-Gow et al., 2004). The binding of CCR3 ligands to CCR3 on the surface of eosinophils causes a transient increase in intracellular Ca 2ϩ concentration ([Ca 2ϩ ] i ) that activates several signal pathways, including actin polymerization (Elsner et al., 1996). With activation of the intracellular motile apparatus, eosinophils undergo shape change

show abstract

Structure Function Differences in Nonpeptide CCR1 Antagonists for Human and Mouse CCR1

Cited by 27 publications

References 20 publications

Predictions of CCR1 Chemokine Receptor Structure and BX 471 Antagonist Binding Followed by Experimental Validation

Predictions of CCR1 Chemokine Receptor Structure and BX 471 Antagonist Binding Followed by Experimental Validation

Site-directed Mutagenesis of CC Chemokine Receptor 1 Reveals the Mechanism of Action of UCB 35625, a Small Molecule Chemokine Receptor Antagonist

A Novel, Selective, and Orally Available Antagonist for CC Chemokine Receptor 3

Contact Info

Product

Resources

About