A Rational Utilization of High-Throughput Screening Affords Selective, Orally Bioavailable 1-Benzyl-3-carboxyazetidine Sphingosine-1-phosphate-1 Receptor Agonists

Hale, Jeffrey J.; Lynch, Christopher L.; Neway, William E.; Mills, Sander G.; Hajdu, Richard; Keohane, Carol A.; Rosenbach, Mark; Milligan, James A.; Shei, Gan-Ju; Parent, Stephen A.; Chrebet, Gary; Bergstrom, James D.; Card, Deborah; Ferrer, Marc; Hodder, Peter; Strulovici, Berta; Rosen, Hugh; Mandala, Suzanne

doi:10.1021/jm0492507

Cited by 84 publications

(72 citation statements)

References 11 publications

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“…Binding assays carried out by Hale et al 37 show S1P 2 does not bind FTY720P, phosphonate and azetidine. In contrast to experimental studies, docking results with the S1P 2 receptor model indicated this receptor interacts favorably with all studied ligands, except SEW2871 ( Figure 5A).…”

Section: Strengths/weaknesses Of the S1p 2 Receptor Modelmentioning

confidence: 98%

Molecular recognition in the sphingosine 1-phosphate receptor family

Pham

Fells

Osborne

et al. 2008

Journal of Molecular Graphics and Modelling

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Computational modeling and its application in ligand screening and ligand receptor interaction studies play important roles in structure-based drug design. A series of sphingosine 1-phosphate (S1P) receptor ligands with varying potencies and receptor selectivities were docked into homology models of the S1P 1-5 receptors. These studies provided molecular insights into pharmacological trends both across the receptor family as well as at single receptors. This study identifies ligand recognition features that generalize across the S1P receptor family, features unique to the S1P 4 and S1P 5 receptors, and suggests significant structural differences of the S1P 2 receptor. Docking results reveal a previously unknown sulfur-aromatic interaction between the S1P 4 C5.44 sulfur atom and the phenyl ring of benzimidazole as well as π-π interaction between F3.33 of S1P 1,4,5 and aromatic ligands. The findings not only confirm the importance of a cation-π interaction between W4.64 and the ammonium of S1P at S1P 4 but also predict the same interaction at S1P 5 . S1P receptor models are validated for pharmacophore development including database mining and new ligand discovery and serve as tools for ligand optimization to improve potency and selectivity.

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Section: Strengths/weaknesses Of the S1p 2 Receptor Modelmentioning

confidence: 98%

Molecular recognition in the sphingosine 1-phosphate receptor family

Pham

Fells

Osborne

et al. 2008

Journal of Molecular Graphics and Modelling

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“…11-13 The S1P 1 receptor is the target of a novel immunosuppressive agent in phase III clinical trials to treat transplant rejection 14 and is the focus of ongoing efforts in multiple laboratories to identify novel agonists with similar therapeutic promise. [15][16][17][18][19][20][21][22][23][24] GPCR exhibit conformational equilibrium between active and inactive conformations. 25,26 In the simplest model of ligand influence on GPCR equilibria, ligands can bind to and stabilize the active conformation (agonist), the inactive conformation (inverse agonist) or can bind to both conformations without preference (neutral antagonist).…”

Section: Introductionmentioning

confidence: 99%

Sphingosine 1-phosphate pKa and binding constants: Intramolecular and intermolecular influences

Naor

Walker

Brocklyn

et al. 2007

Journal of Molecular Graphics and Modelling

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The dissociation constant for an ionizable ligand binding to a receptor is dependent on its charge and therefore on its environmentally-influenced pK a value. The pK a values of sphingosine 1-phosphate (S1P) were studied computationally in the context of the wild type S1P 1 receptor and the following mutants: E3.29Q, E3.29A, and K5.38A. Calculated pK a values indicate that S1P binds to S1P 1 and its site mutants with a total charge of −1, including a +1 charge on the ammonium group and a −2 charge on the phosphate group. The dissociation constant of S1P binding to these receptors was studied as well. The models of wild type and mutant proteins originated from an active receptor model that was developed previously. We used ab initio RHF/6-31+G(d) to optimize our models in aqueous solution, where the solvation energy derivatives are represented by conductor-like polarizable continuum model (C-PCM) and integral equation formalism polarizable continuum model (IEF-PCM). Calculation of the dissociation constant for each mutant was determined by reference to the experimental dissociation constant of the wild type receptor. The computed dissociation constants of the E3.29Q and E3.29A mutants are 3-5 orders of magnitude higher than those for the wild type receptor and K5.38A mutant, indicating vital contacts between the S1P phosphate group and the carboxylate group of E3.29. Computational dissociation constants for K5.38A, E3.29A and E3.29Q mutants were compared with experimentally determined binding and activation data. No measurable binding of S1P to the E3.29A and E3.29Q mutants was observed, supporting the critical contacts observed computationally. These results validate the quantitative accuracy of the model.

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“…Fluorescence intensity was measured using the LSRII FACS machine (Becton Dickinson, Franklin Lakes, NJ). A previously published compound, 1-(3-methyl-4-((4-pheyl-5-(trifluoromethyl)thiophen-2-yl)methoxy)benzyl)zaetidine-3-carboxylic acid, 32 at 100 nM was used as the low control (internalized S1P 1 ), and the value of DMSO only was used as the high control (S1P 1 on surface). Internalization results were captured as the geometric mean of the mean fluorescence intensity.…”

Section: Receptor Internalizationmentioning

confidence: 99%

Predictability of Peripheral Lymphocyte Reduction of Novel S1P1 Agonists by In Vitro GPCR Signaling Profile

McElvain

Fiorino

et al. 2013

SLAS Discovery

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Surrogate readouts of G-protein-coupled receptor signaling pathways using highly engineered systems are often employed in the drug discovery process. However, accumulating data have demonstrated the importance of selecting relevant biological activity rather than technically facile assays to support high-throughout screening and subsequent structureactivity relationship studies. Here we report a case study using sphingosine-1-phosphate receptor 1 (S1P 1 ) as the model system to compare compound activity in six different in vitro assays with their ability to predict in vivo efficacy. S1P 1 has long been validated as a therapeutic target for autoimmune diseases. In this article, in vivo and in vitro studies on 19 S1P 1 agonists are reported. In vitro activities of these S1P 1 agonists, together with S1P and FTY720p, on Ca 2+ mobilization, adenylyl cyclase inhibition, extracellular signal-related kinase (ERK) phosphorylation, β-arrestin recruitment, and receptor internalization, were determined. The in vitro potency of these compounds was correlated with their ability to induce peripheral lymphocyte reduction. The results revealed that inhibition of adenylyl cyclase and induction of β-arrestin recruitment and receptor internalization are good indicators to predict in vivo efficacy, whereas induction of Ca 2+ mobilization through G qi/5coupling and ERK phosphorylation is irrelevant. This study demonstrated the importance of identifying an appropriate in vitro assay to predict in vivo activity based on the biological relevance in the drug discovery setting.

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A Rational Utilization of High-Throughput Screening Affords Selective, Orally Bioavailable 1-Benzyl-3-carboxyazetidine Sphingosine-1-phosphate-1 Receptor Agonists

Cited by 84 publications

References 11 publications

Molecular recognition in the sphingosine 1-phosphate receptor family

Molecular recognition in the sphingosine 1-phosphate receptor family

Sphingosine 1-phosphate pKa and binding constants: Intramolecular and intermolecular influences

Predictability of Peripheral Lymphocyte Reduction of Novel S1P1 Agonists by In Vitro GPCR Signaling Profile

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