Discovery of 2-(Phenoxypyridine)-3-phenylureas as Small Molecule P2Y<sub>1</sub> Antagonists

Chao, Hannguang J.; Turdi, Huji; Herpin, Timothy F.; Roberge, Jacques Y.; Liu, Yalei; Schnur, Dora M.; Poss, Michael A.; Rehfuss, Robert; Hua, Jinping; Wu, Qimin; Price, Laura A.; Abell, Lynn M.; Schumacher, William A.; Bostwick, Jeffrey S.; Steinbacher, Thomas E.; Stewart, Anne B.; Ogletree, Martin L.; Huang, Christine; Chang, Ming; Cacace, Angela; Arcuri, Maredith; Celani, Deborah; Wexler, Ruth R.; Lawrence, R. Michael

doi:10.1021/jm301708u

Cited by 50 publications

(47 citation statements)

References 34 publications

(26 reference statements)

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“…However, it should be noted that only very few allosteric antagonists have been used in crystallization compared with the total population of GPCR antagonists available. Even for the urea class of P2Y 1 R antagonists related to BPTU, the binding location was initially predicated by molecular modeling to be within the helical bundle (Chao et al, 2013;Qiao et al, 2013). Thus, potentially biased functional antagonism remains to be explored for other P2Y 1 R antagonists and for diverse antagonists, especially allosteric modulators, of other GPCRs.…”

Section: Discussionmentioning

confidence: 99%

“…1) bound in a pocket formed within the seven transmembrane (TM) domains and more external than most small ligands of rhodopsin-like GPCRs. However, an allosteric antagonist 1-(2-(2-(tert-butyl)phenoxy)-pyridin-3-yl)-3-(4-(trifluoromethoxy)phenyl)urea (BPTU), a hydrophobic diaryl-urea derivative that arose from a program to develop antithrombotic drugs (Chao et al, 2013), bound on the external receptor surface at the phospholipid membrane interface in contact with TMs 1-3 (Zhang et al, 2015a). This represented the first example in the GPCR field of a ligand located outside the helical bundle or loop regions.…”

Section: Introductionmentioning

confidence: 99%

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Distinct Signaling Patterns of Allosteric Antagonism at the P2Y₁ Receptor

Gao

2017

Mol Pharmacol

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Traditionally, G protein-coupled receptor antagonists are classified as competitive or noncompetitive and surmountable or insurmountable based on functional antagonism. P2Y 1 receptor (P2Y 1 R) structures showed two antagonists binding to two spatially distinct sites: nucleotide MRS2500 (orthosteric, contacting the helical bundle) and urea BPTU (allosteric, on the external receptor surface). However, the nature of their P2Y 1 R antagonism has not been characterized. Here we characterized BPTU antagonism at various signaling pathways activated by structurally diverse agonists. BPTU rightward shifted the concentration-response curves of both 2-methylthioadenosine 59-diphosphate trisodium salt and MRS2365 (59-diphosphates) in some signaling events, such as extracellular signal-regulated kinase 1/2 and label free, in a parallel manner without affecting the maximum agonist effect (E max ) but antagonized insurmountably (suppressed agonist E max ) in signaling events such as guanosine 59-3-O-(thio)triphosphate binding and b-arrestin2 recruitment. However, with dinucleotide Ap4A as an agonist, BPTU suppressed the E max insurmountably in all signaling pathways. By comparison, MRS2500 behaved as surmountable antagonist rightward-shifting concentration-response curves of all three agonists in a parallel manner for all signaling pathways measured. Thus, we demonstrated a previously undocumented phenomenon that P2Y 1 R antagonism patterns could vary in different signaling pathways, which could be related to conformational selection, signaling amplification, and probe dependence. This phenomenon may apply generally to other receptors considering that antagonism by a specific ligand is often not compared at multiple signaling pathways. Thus, antagonism can be surmountable or insurmountable depending on the signaling pathways measured and the agonists used, which should be of broad relevance to drug discovery and disease treatment.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Distinct Signaling Patterns of Allosteric Antagonism at the P2Y₁ Receptor

Gao

2017

Mol Pharmacol

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“…Semitransparent surface of binding site's residues is displayed in pale cyan. (Chao et al, 2013) were recently determined (Fig. 4B) (Zhang et al, 2015).…”

Section: Structural Characterization Of P2yrsmentioning

confidence: 94%

Nucleotides Acting at P2Y Receptors: Connecting Structure and Function

et al. 2015

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Eight G protein-coupled P2Y receptor (P2YR) subtypes are important physiologic mediators. The human P2YRs are fully activated by ATP (P2Y 2 and P2Y 11 ), ADP (P2Y 1 , P2Y 12 , and P2Y 13 ), UTP (P2Y 2 and P2Y 4 ), UDP (P2Y 6 and P2Y 14 ), and UDP glucose (P2Y 14 ). Their structural elucidation is progressing rapidly. The X-ray structures of three ligand complexes of the G i -coupled P2Y 12 R and two of the G q -coupled P2Y 1 Rs were recently determined and will be especially useful in structure-based ligand design at two P2YR subfamilies. These high-resolution structures, which display unusual binding site features, complement mutagenesis studies for probing ligand recognition and activation. The structural requirements for nucleotide agonist recognition at P2YRs are relatively permissive with respect to the length of the phosphate moiety, but less so with respect to base recognition. Nucleotide-like antagonists and partial agonists are also known for P2Y 1 , P2Y 2 , P2Y 4 , and P2Y 12 Rs. Each P2YR subtype has the ability to be activated by structurally bifunctional agonists, such as dinucleotides, typically, dinucleoside triphosphates or tetraphosphates, and nucleoside polyphosphate sugars (e.g., UDP glucose) as well as the more conventional mononucleotide agonists. A range of dinucleoside polyphosphates, from triphosphates to higher homologs, occurs naturally. Earlier modeling predictions of the P2YRs were not very accurate, but recent findings have provided much detailed structural insight into this receptor family to aid in the rational design of new drugs.

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“…In their discovery program a high throughput screening (HTS) effort, using >1 million compounds against the human P2Y1R, revealed that diarylurea 8 30 had good affinity and binding selectivity toward the P2Y1R versus P2Y12R (P2Y1 Ki = 75 nM, P2Y12 Ki > 70 μM).…”

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confidence: 99%

Drug-like Antagonists of P2Y Receptors—From Lead Identification to Drug Development

et al. 2016

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P2Y receptors are expressed in virtually all cells and tissue types and mediate an astonishing array of biological functions including platelet aggregation, smooth muscle cell proliferation and immune regulation. The P2Y receptors belong to the GPCR superfamily and are composed of eight members encoded by distinct genes that can be subdivided into two groups based on their coupling to specific G-proteins. Extensive research has been undertaken to find modulators of P2Y receptors, although to date there have only been a limited number of small molecule P2Y receptor antagonists that have been approved by drug/medicines agencies. This Perspective reviews the known P2Y receptor antagonists, highlighting oral drug-like receptor antagonists, and considers future opportunities for the development of small molecules for clinical evaluation.

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Discovery of 2-(Phenoxypyridine)-3-phenylureas as Small Molecule P2Y₁ Antagonists

Cited by 50 publications

References 34 publications

Distinct Signaling Patterns of Allosteric Antagonism at the P2Y₁ Receptor

Distinct Signaling Patterns of Allosteric Antagonism at the P2Y₁ Receptor

Nucleotides Acting at P2Y Receptors: Connecting Structure and Function

Drug-like Antagonists of P2Y Receptors—From Lead Identification to Drug Development

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Discovery of 2-(Phenoxypyridine)-3-phenylureas as Small Molecule P2Y1 Antagonists

Cited by 50 publications

References 34 publications

Distinct Signaling Patterns of Allosteric Antagonism at the P2Y1 Receptor

Distinct Signaling Patterns of Allosteric Antagonism at the P2Y1 Receptor

Nucleotides Acting at P2Y Receptors: Connecting Structure and Function

Drug-like Antagonists of P2Y Receptors—From Lead Identification to Drug Development

Contact Info

Product

Resources

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Discovery of 2-(Phenoxypyridine)-3-phenylureas as Small Molecule P2Y₁ Antagonists

Distinct Signaling Patterns of Allosteric Antagonism at the P2Y₁ Receptor

Distinct Signaling Patterns of Allosteric Antagonism at the P2Y₁ Receptor