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Prostaglandin (PG) D2 elicits responses through either the DP1 and/or DP2 receptor. Experimental evidence suggests that stimulation of the DP1 receptor contributes to allergic responses, such that antagonists are considered to be directed therapies for allergic diseases. In this study, we demonstrate the activity of a novel synthetic DP1 receptor antagonist termed asapiprant (S-555739) for the DP1 receptor and other receptors in vitro, and assess the efficacy of asapiprant in several animal models of allergic diseases. We determined the affinity and selectivity of asapiprant for the DP1 receptor in binding assays. In the animal models of allergic rhinitis, changes in nasal resistance, nasal secretion, and cell infiltration in nasal mucosa were assessed after antigen challenge with and without asapiprant. Similarly, in the animal models of asthma, the effect of antigen challenge with and without asapiprant on antigen-induced bronchoconstriction, airway hyper-responsiveness, mucin production, and cell infiltration in lung were assessed. In binding studies, asapiprant exhibited high affinity and selectivity for the DP1 receptor. Significant suppression of antigen-induced nasal resistance, nasal secretion, and cell infiltration in nasal mucosa was observed with asapiprant treatment. In addition, treatment with asapiprant suppressed antigen-induced asthmatic responses, airway hyper-responsiveness, and cell infiltration and mucin production in lung. These results show that asapiprant is a potent and selective DP1 receptor antagonist, and exerts suppressive effects in the animal models of allergic diseases. Thus, asapiprant has potential as a novel therapy for allergic airway diseases., (Copyright © 2015 Elsevier B.V. All rights reserved.)

The chemoattractant receptor-homologous molecule expressed on Th2 cells (CRTH2) is a G protein-coupled receptor that mediates the pro-inflammatory effects of prostaglandin D(2) (PGD(2)) generated in allergic inflammation. The CRTH2 receptor shares greatest sequence similarity with chemoattractant receptors compared with prostanoid receptors. To investigate the structural determinants of CRTH2 ligand binding, we performed site-directed mutagenesis of putative mCRTH2 ligand-binding residues, and we evaluated mutant receptor ligand binding and functional properties. Substitution of alanine at each of three residues in the transmembrane (TM) helical domains (His-106, TM III; Lys-209, TM V; and Glu-268, TM VI) and one in extracellular loop II (Arg-178) decreased PGD(2) binding affinity, suggesting that these residues play a role in binding PGD(2). In contrast, the H106A and E268A mutants bound indomethacin, a nonsteroidal anti-inflammatory drug, with an affinity similar to the wild-type receptor. HEK293 cells expressing the H106A, K209A, and E268A mutants displayed reduced inhibition of intracellular cAMP and chemotaxis in response to PGD(2), whereas the H106A and E268A mutants had functional responses to indomethacin similar to the wild-type receptor. Binding of PGE(2) by the E268A mutant was enhanced compared with the wild-type receptor, suggesting that Glu-268 plays a role in determining prostanoid ligand selectivity. Replacement of Tyr-261 with phenylalanine did not affect PGD(2) binding but decreased the binding affinity for indomethacin. These results provided the first details of the ligand binding pocket of an eicosanoid-binding chemoattractant receptor.

A novel series of 15-fluoro prostaglandins with phenoxy termination of the omega-chain was synthesized and evaluated for binding and functional activation of the prostaglandin FP receptor in vitro and for side effect potential and topical ocular hypotensive efficacy in vivo. Compounds with the 15alpha-fluoride relative stereochemistry displayed EC50 values of

Prostanoids are a group of lipid mediators that include the prostaglandins (PG) and thromboxanes (TX). Upon cell stimulation, prostanoids are synthesized from arachidonic acid via the cyclooxygenase (COX) pathway and released outside the cells to exert various physiological and pathological actions in a variety of tissues and cells. The activities of prostanoids are mediated by specific G protein-coupled receptors, which have been classified on the basis of pharmacological experiments into eight types and subtypes according to their responsiveness to selective agonists and antagonists. These prostanoid receptors have been cloned from various species including human, and their distinct binding properties and signal transduction pathways have been characterized by analyses of cells expressing each receptor. Furthermore, the distribution patterns of prostanoid receptor mRNAs have been determined in tissues and cells for various species. This information is useful for understanding the molecular basis of the pathophysiological actions of prostanoids.

Cyclopentenone prostaglandins (PGs), such as 15-deoxy-12,13-didehydro-14,15-didehydro-PGJ2 (15d-delta(12,14)-PDJ2), 12,13-didehydro-PGJ2 (delta12-PGJ2) and PGA2, are actively transported into cells and promote the expression of a variety of genes. The ultimate metabolite of PGD2, 15d-delta(12,14)-PGJ2, specifically binds to a nuclear receptor, the gamma isoform of the peroxisome proliferator-activated receptor, thereby promoting adipogenesis. Cyclopentenone PGs also induce the expression of various stress genes, such as heat shock proteins (HSPs), the immunoglobulin heavy chain binding protein (BiP) and protein disulfide isomerase by acting through heat shock element or unfolded protein response element. Overall, cyclopentenone PGs regulate cell growth, cell differentiation and stress responses by regulating various gene expression.

The in vitro evaluation of a series of saturated prostaglandins revealed that compounds with omega chain aromatic rings retain nanomolar potency for the human prostaglandin F receptor (hFP receptor), exemplified by compound 8. In contrast, the double bonds are required for activity in the series with an acyclic omega chain as in PGF2alpha.

Prostanoid receptors are the G-protein-coupled, rhodopsin-type receptors with seven transmembrane domains and consist of eight types and subtypes. Although the overall homology is not high, there are several regions specifically conserved among them. These regions are considered to form the ligand binding pocket for the structures common to prostanoid molecules, and the other regions to confer specificity for ligand binding. The PGI and PGD receptors have relatively high homology (40%) at the amino acid level and share the same signalling pathway. To determine which structural domains of these receptors confer ligand binding specificity, we constructed a series of chimeric receptors from the mouse PGI and PGD receptors. These chimeric receptors were expressed in COS-7 cells, and their abilities to bind prostaglandins and their analogues were examined. The region from the sixth transmembrane domain to the carboxyl terminus of the PGI receptor was first replaced by the corresponding region of the PGD receptor. This chimeric receptor binds both PGD2 and PGE2, though the ability to bind iloprost, a PGI receptor agonist, and PGE1 does not change. This result indicates that the sixth and seventh transmembrane domains of the PGI receptor play an important role in distinction of structural difference between PGE1 and PGE2 in the alpha-side chain. These binding characteristics did not change when the region up to the third transmembrane domain of the PGI receptor was replaced with the corresponding region of the PGD receptor. However, when the first extracellular loop including a portion of the second transmembrane domain was further replaced, the abilities to bind PGE1, PGE2 and iloprost were eliminated. This result indicates that this domain of the PGD receptor is responsible for distinction of structural differences between PGD2 and PGE2 on the cyclopentane ring.