P2Y14 receptor

Aims: UDP-sugars can act as extracellular signalling molecules, but relatively little is known about their cardiovascular actions. The P2Y14 receptor is a Gi/o-coupled receptor which is activated by UDP-glucose and related sugar nucleotides. In this study we sought to investigate whether P2Y14 receptors are functionally expressed in the porcine coronary artery using a selective P2Y14 receptor agonist, MRS2690, and a novel selective P2Y14 receptor antagonist, PPTN (4,7-disubstituted naphthoic acid derivative). Methods and results: Isometric tension recordings were used to evaluate the effects of UDP-sugars in porcine isolated coronary artery segments. The effects of the P2 receptor antagonists suramin and PPADS, the P2Y14 receptor antagonist PPTN, and the P2Y6 receptor antagonist MRS2578, were investigated. Measurement of vasodilator-stimulated phosphoprotein (VASP) phosphorylation using flow cytometry was used to assess changes in cAMP levels. UDP-glucose, UDP-glucuronic acid UDP-N-acetylglucosamine (P2Y14 receptor agonists), elicited concentration-dependent contractions of the porcine coronary artery. MRS2690 was a more potent vasoconstrictor than the UDP-sugars. Concentration dependent contractile responses to MRS2690 and UDP-sugars were enhanced in the presence of forskolin (activator of cAMP), where the level of basal tone was maintained by addition of U46619, a thromboxane A2 mimetic. Contractile responses to MRS2690 were blocked by PPTN, but not by MRS2578. Contractile responses to UDP-glucose were also attenuated by PPTN and suramin, but not by MRS2578. Forskolin-induced VASP-phosphorylation was reduced in porcine coronary arteries exposed to UDP-glucose and MRS2690, consistent with P2Y14 receptor coupling to Gi/o proteins and inhibition of adenylyl cyclase activity. Conclusions: Our data support a role of UDP-sugars as extracellular signalling molecules and show for the first time that they mediate contraction of porcine coronary arteries via P2Y14 receptors [1].

[1]. UDP-sugars activate P2Y14 receptors to mediate vasoconstriction of the porcine coronary artery. Vascul Pharmacol. 2018 Apr:103-105:36-46.

Background: Enterochromaffin cells (ECs) synthesize and release serotonin (5-HT) and adenosine triphosphate (ATP) to trigger or regulate enteroneurinic reflexes and transmit visceral/pain sensory information. Alterations in the 5-HT signaling pathway may be involved in the pathogenesis of inflammatory bowel disease (IBD) or irritable bowel syndrome (IBS), but the pharmacological or molecular mechanisms regulating Ca2+-dependent 5-HT release remain unclear. Previous studies have shown that purinergic signaling via ATP and ADP is an important mechanism regulating 5-HT release. However, ECs also respond to uridine triphosphate (UTP) and uridine diphosphate (UDP), suggesting that uridine triphosphate receptors and signaling pathways are also involved. We tested the hypothesis that UTP is a regulator of 5-HT release in human ECs. Methods: We investigated the UTP signaling pathway mechanism in human EC model BON cells using Fluo-4/Ca2+ imaging, patch-clamp, pharmacological analysis, immunohistochemistry, Western blotting, and qPCR. The release of serotonin (5-HT) was monitored in enteric ganglion cells (hECs) or osteochondral cells (BONs) isolated from human intestinal surgical specimens. Results: UTP, UTPγS, UDP, and ATP all induced Ca2+ oscillations in BONs. UTP induced a concentration-dependent biphasic Ca2+ response. The cellular response order was: UTP, ATP > UTPγS > UDP >> MRS2768, BzATP, α,β-MeATP > MRS2365, MRS2690, and NF546. Cells activated by UTP and ATP at varying proportions also responded to UTPγS (P2Y4, 50% of cells), UDP (P2Y6, 30%), UTPγS and UDP (14%), or MRS2768 (<3%). UTP-induced Ca2+ responses could be blocked by PLC, IP3R, SERCA Ca2+ pumps, inhibitors of La3+-sensitive Ca2+ channels, or BAPTA/AM chelation of intracellular free Ca2+. Inhibitors of L-type calcium channels, TRPC, raneidin-Ca2+ pools, PI3 kinase, PKC, or SRC kinases had no such effect. UTP stimulated depolarization of voltage-sensitive Ca2+ current (ICa) and membrane potential (Vm), and inhibited IK current (but not IA current). The IKv7.2/7.3 K+ channel blocker XE-991 mimicked UTP-induced Vm depolarization and blocked the UTP response. XE-991 blocks IK currents, while UTP further reduces them. La3+ or PLC inhibitors block UTP-induced depolarization; PKC inhibitors, thapsigargin, or zero Ca2+ buffer have no such effect. UTP stimulates the release of 5-HT from hEC cells expressing TPH1, 5-HT, and P2Y4/P2Y6R. Zero Ca2+ buffer enhances Ca2+ response and 5-HT release. Conclusion: UTP activates the major P2Y4R pathway, mobilizing intracellular Ca2+ via the PLC/IP3/IP3R/SERCA Ca2+ signaling pathway, thereby triggering Ca2+ oscillations and stimulating 5-HT release; Ca2+ influx is inhibitory. UTP-induced membrane potential depolarization depends on the PLC signaling pathway and an unidentified K channel (which appears to be independent of Ca2+ oscillations or Ica/VOCC). A UTP-gated signaling pathway triggered by P2Y4R activation stimulates 5-HT release. Front Pharmacol. 2017 Jul 13:8:429.

C15H22N2NA2O16P2S

626.32

625.996

C, 28.77; H, 3.54; N, 4.47; Na, 7.34; O, 40.87; P, 9.89; S, 5.12

15039-58-4

73755042

Typically exists as solid at room temperature

7

17

9

38

981

9

[Na+].[Na+].OC[C@H]1O[C@H](OP(OP(OC[C@H]2O[C@@H](N3C=CC(=O)NC3=S)[C@H](O)[C@@H]2O)(=O)[O-])(=O)[O-])[C@H](O)[C@@H](O)[C@@H]1O

TYVFMVSNSGMZPA-QBNUFUENSA-L

InChI=1S/C15H24N2O16P2S.2Na/c18-3-5-8(20)10(22)12(24)14(31-5)32-35(27,28)33-34(25,26)29-4-6-9(21)11(23)13(30-6)17-2-1-7(19)16-15(17)36;;/h1-2,5-6,8-14,18,20-24H,3-4H2,(H,25,26)(H,27,28)(H,16,19,36);;/q;2*+1/p-2/t5-,6-,8-,9-,10+,11-,12-,13-,14-;;/m1../s1

disodium;[[(2R,3S,4R,5R)-3,4-dihydroxy-5-(4-oxo-2-sulfanylidenepyrimidin-1-yl)oxolan-2-yl]methoxy-oxidophosphoryl] [(2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl] phosphate

MRS 2690; MRS-2690; MRS2690; MRS 2690; MRS-2690; 15039-58-4; GTPL3337; GLXC-04134; [[[(2R,3S,4R,5R)-3,4-dihydroxy-5-(4-oxo-2-sulfanylidenepyrimidin-1-yl)oxolan-2-yl]methoxy-sodiooxyphosphoryl]oxy-[(2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyphosphoryl]oxysodium; MRS 2690; DIPHOSPHORIC ACID 1-A-D-GLUCOPYRANOSYL ESTER 2-[(4'-METHYLTHIO)URIDIN-5''-YL] ESTER DISODIUM SALT; MRS2690

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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 (Please use freshly prepared in vivo formulations for optimal results.)