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| Targets |
Ki: 0.6 nM (human EP1), 1.7 nM (mouse EP1)[1]
ONO-8711 dicyclohexylamine specifically targets the EP1 receptor, one of four G protein-coupled receptors (GPCRs) that mediate the diverse physiological and pathophysiological effects of prostaglandin E2 (PGE2). Unlike EP2, EP3, and EP4 receptors, which primarily couple to Gs and Gi proteins, the EP1 receptor couples to Gq. Activation of EP1 by PGE2 leads to an increase in intracellular calcium (Ca2+) via phospholipase C (PLC) activation. This calcium signaling triggers smooth muscle contraction, neuronal activation, and pain transmission. By acting as a competitive antagonist, ONO-8711 binds to the EP1 receptor and blocks PGE2 from binding, thereby inhibiting EP1-mediated calcium mobilization and downstream cellular responses. It is used to study EP1's role in various pathological conditions. |
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| ln Vitro |
ONO-8711 (10 and 30 μM; 30 min) non-competitively inhibits sulprostone-induced pulmonary vein contractions in humans[2]. For the rat, mouse, and human receptors, ONO-8711 has IC50s of 0.22 μM, 0.05 μM, and 0.21 μM, respectively, and suppresses the PGE2-induced rise in cytosolic Ca2+ concentration[3].
In vitro, ONO-8711 dicyclohexylamine potently inhibits PGE2 binding to EP1 receptors. In receptor binding assays using membranes from CHO cells expressing human EP1, the compound exhibits a Ki of 0.6 nM. In membranes expressing mouse EP1, the Ki is 1.7 nM. The selectivity profile has been characterized against a panel of prostanoid receptors: Ki for mouse EP2 > 1,000 nM; Ki for mouse EP3 = 67 nM; Ki for mouse EP4 > 1,000 nM; and Ki for mouse TP > 1,000 nM. Functional antagonism is demonstrated by inhibiting PGE2-induced calcium mobilization in EP1-expressing cells. In CHO cells expressing human EP1, ONO-8711 (0.1-100 nM) blocks PGE2 (10 nM)-induced intracellular calcium increase, as measured using Fluo-4 AM fluorescence. The IC50 for functional antagonism is in the low nanomolar range, consistent with the binding affinity. |
| ln Vivo |
ONO-8711 (400 or 800 ppm; po; for 20 weeks) inhibits the growth of breast tumors and reduces the incidence of cancer[3].
The in vivo activity of ONO-8711 dicyclohexylamine has been studied in animal models of inflammation and pain. As an orally active compound, ONO-8711 demonstrates efficacy in models where EP1 activation contributes to disease pathology. In rodent models of inflammatory pain (e.g., carrageenan-induced mechanical hyperalgesia), oral administration of ONO-8711 (0.1-10 mg/kg) reverses PGE2-induced hyperalgesia. In models of colitis and bladder inflammation, EP1 antagonists reduce visceral pain. ONO-8711 has also been used to study EP1-mediated signaling in cancer models, such as colon cancer and breast cancer, where EP1 expression correlates with tumor progression. The compound is orally bioavailable and has a favorable pharmacokinetic profile for in vivo research. |
| Enzyme Assay |
Cell-free receptor binding assays are performed using membrane preparations from CHO cells stably expressing human or mouse EP1 receptors. Membranes (10-20 microg protein/well) are incubated with the radioligand [3H]-PGE2 (1 nM) and varying concentrations of ONO-8711 dicyclohexylamine (0.01 pM to 10 microM) in binding buffer (50 mM Tris-HCl, pH 6.0, 10 mM MgCl2, 1 mM EDTA, 0.1% BSA) for 60-90 minutes at 25degC. Non-specific binding is determined by adding 10 microM unlabeled PGE2. The reaction is terminated by rapid vacuum filtration through GF/B glass fiber filters presoaked in 0.5% polyethylenimine (PEI). Filters are washed three times with ice-cold buffer, dried, and counted. Ki values are calculated from the IC50 using the Cheng-Prusoff equation. The compound exhibits Ki = 0.6 nM for human EP1 and Ki = 1.7 nM for mouse EP1. Selectivity is assessed by similar binding assays against EP2, EP3, EP4, TP, and FP.
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| Cell Assay |
A functional calcium mobilization assay is performed using CHO-K1 cells stably expressing human EP1 receptors. Cells are seeded in 96-well black-walled clear-bottom plates (3×10⁴ cells/well) and cultured overnight. Cells are loaded with 2 microM Fluo-4 AM and 0.02% Pluronic F-127 in HBSS (with Ca2+ and Mg2+) for 60 minutes at 37degC, followed by a 30-minute de-esterification period at room temperature. Various concentrations of ONO-8711 dicyclohexylamine (0.01-1000 nM) are added and incubated for 15-30 minutes to block the EP1 receptor. Then, PGE2 (10 nM, EC80 concentration) is added to the cells, and the fluorescence signal (ex/em 494/516 nm) is measured immediately for 1-2 minutes using a fluorescence plate reader (e.g., FlexStation 3). Peak fluorescence increase is recorded. The percent inhibition of the PGE2-induced calcium signal is calculated relative to the vehicle control. The IC50 is determined from the dose-response curve. A Schild analysis can be performed to confirm competitive antagonism.
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| Animal Protocol |
Animal/Disease Models: Female SD (Sprague-Dawley) rats (induced breast cancer by gavage of 85 mg/kg PhIP (HY -118716) 4 times for 2 weeks)
Doses: 400 or 800 ppm Route of Administration: po; for 20 weeks Experimental Results: Did not induce any symptoms of toxicity at 800 ppm. Delayed occurrence of breast tumors for 2 or 4 weeks at 400 or 800 ppm , respectively. Dramatically suppressed cancer incidence compared with the control diet group at 800 ppm (56% versus 79%, P < 0.05). The in vivo efficacy of ONO-8711 dicyclohexylamine can be evaluated in a carrageenan-induced mechanical hyperalgesia model in rats. Male Sprague-Dawley rats (200-250 g) are used. On the test day, baseline paw withdrawal thresholds (PWT) are measured using an electronic von Frey anesthesiometer. ONO-8711 is formulated in an appropriate vehicle (e.g., 0.5% methylcellulose or 10% DMSO, 5% Tween-80, 85% saline) and administered orally at doses of 0.1, 0.3, 1, 3, and 10 mg/kg. One hour after oral administration (or at the Tmax determined from PK studies), 50 microL of 1% carrageenan is injected intraplantarly into the hind paw. Mechanical hyperalgesia (reduction in paw withdrawal threshold) is measured 3, 4, and 5 hours after carrageenan injection. The area under the time-PWT curve (AUC) is calculated. The ED50 for reversal of hyperalgesia is determined. A separate group receives PGE2 (100 ng) directly into the paw (intraplantar) instead of carrageenan. In this model, ONO-8711 blocks PGE2-induced hyperalgesia, confirming EP1 specificity. Body weight is monitored as a toxicity indicator. |
| ADME/Pharmacokinetics |
ONO-8711 dicyclohexylamine has a molecular weight of 596.84 g/mol (free base). The compound is supplied as a solid and should be stored at -20degC in a sealed container, protected from light and moisture, where it is stable for up to 3 years. For solution storage, it should be kept at -80degC for up to 6 months. The compound is soluble in DMSO (e.g., 10-20 mg/mL) and ethanol. For oral administration, it can be formulated in 0.5% methylcellulose or 10% DMSO + 5% Tween-80 + 85% saline. The dicyclohexylamine salt is used to improve the solubility and handling properties of the parent free base. The compound is a competitive EP1 antagonist. The product is for research use only, not for human consumption.
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| Toxicity/Toxicokinetics |
This product is for research use only and is not for human therapeutic use. No specific toxicity data is available for this compound. In preclinical studies in rodents, ONO-8711 dicyclohexylamine was generally well-tolerated at doses up to 30 mg/kg orally. No significant acute toxicity or body weight loss was reported at the effective doses (0.1-10 mg/kg) used for in vivo pharmacology studies. The EP1 receptor is expressed in various tissues, including the kidneys, bladder, uterus, and dorsal root ganglia. Its inhibition may affect physiological processes such as blood pressure regulation, sodium excretion, and parturition. Standard laboratory safety practices (gloves, lab coat, safety glasses) should be followed. Avoid inhalation of powder and contact with skin. The compound is not an FDA-approved drug.
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| References |
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| Additional Infomation |
Prostaglandin E2 (PGE2) is a major pro-inflammatory lipid mediator that acts through four receptors: EP1, EP2, EP3, and EP4. EP1 is a Gq-coupled receptor that increases intracellular calcium, leading to smooth muscle contraction, neuronal excitation, and pain. EP1 antagonists have been investigated for the treatment of inflammatory pain, overactive bladder, and certain cancers. ONO-8711 is a highly potent and selective EP1 antagonist that has been widely used as a pharmacological tool to elucidate the role of EP1 in various disease models. The compound is structurally distinct and offers excellent selectivity over EP2 and EP4 (>1,000-fold). The dicyclohexylamine salt enhances aqueous solubility. This product is not a clinically approved drug. Supplier information must not be included.
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| Molecular Formula |
C34H53CLN2O4S
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| Molecular Weight |
621.31
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| Related CAS # |
ONO-8711;216158-34-8
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| Appearance |
White to off-white solid powder
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO :~33.33 mg/mL (~53.64 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (4.02 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (4.02 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.6095 mL | 8.0475 mL | 16.0950 mL | |
| 5 mM | 0.3219 mL | 1.6095 mL | 3.2190 mL | |
| 10 mM | 0.1610 mL | 0.8048 mL | 1.6095 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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