| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| Other Sizes |
| Targets |
TNF-α; IL-1β; IL-6; iNOS; NO
GPR4 antagonist (IC₅₀ = 0.070 ± 0.019 µM for human GPR4 in HeLa cells) Selective over related proton-sensing receptors OGR1 and TDAG8 (IC₅₀ > 20 µM) hERG channel binding affinity: 19 µM (44% inhibition at 60 µM in patch-clamp assay) Histamine H₃ receptor binding: >30 µM |
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| ln Vitro |
NE 52-QQ57 efficiently inhibits GPR4-mediated cAMP accumulation (IC50 26.8 nM) in HEK293 cells)[2].
In HeLa cells expressing human GPR4, 13 inhibited pH-induced cAMP release with an IC₅₀ of 0.070 ± 0.019 µM. The compound showed selectivity over OGR1 and TDAG8 (IC₅₀ > 20 µM) and low affinity for the hERG channel (IC₅₀ = 19 µM). In the presence of 40% rat plasma, the IC₅₀ shift for hGPR4 was minimal (factor of 1), indicating low plasma protein binding interference.[1] |
| ln Vivo |
NE 52-QQ57 (Compound 13) demonstrates a notable reduction in inflammation in the rat antigen-induced arthritis model following oral treatment at a dose of 30 mg/kg bid for 20 days[1]. As shown in the rat complete Freund's adjuvant model, NE 52-QQ57 (30 mg/kg bid po for 4 days) also inhibits pain and angiogenesis in the mouse chamber model[1].
In a rat antigen-induced arthritis (AIA) model, 13 administered orally at 30 mg/kg bid for 7 days significantly inhibited knee swelling (34.2 ± 6.6% inhibition). In a mouse VEGF-induced implant angiogenesis chamber model, 13 (30 mg/kg bid po for 4 days) significantly reduced tissue growth by 46.8 ± 10.6%. In a rat complete Freund’s adjuvant (CFA)-induced mechanical hyperalgesia model, 13 showed dose-dependent reversal of hyperalgesia, with maximal reversal of 62 ± 8% and an ED₅₀ of 14.7 mg/kg.[1] |
| Enzyme Assay |
Human GPR4 cAMP release assay: HeLa cells stably expressing human GPR4 were seeded in 384-well plates and incubated with compound in Hepes-buffered saline (pH 8) containing IBMX for 15 min, then stimulated with acidic buffer (pH 6.5) for another 15 min. cAMP levels were measured using HTRF technology.
hERG binding assay: Membranes from HEK293 cells expressing hERG channels were incubated with [³H]dofetilide and compound for 90 min, followed by filtration and scintillation counting. Histamine H₃ receptor binding assay: CHO-K1 cell membranes expressing human H₃ receptors were incubated with [³H]-R-α-methylhistamine and compound in Tris buffer, followed by SPA bead-based detection.[1] |
| Cell Assay |
GPR4 activity was assessed in HeLa cells stably transfected with human, rat, or mouse GPR4. Cells were treated with compound and stimulated with acidic pH, and cAMP production was measured via HTRF.
Selectivity was tested against OGR1 and TDAG8 in similar cAMP assays using transfected HeLa cells.[1] |
| Animal Protocol |
Female FVB mice (8-10 weeks)
30 mg/kg Oral, 4 days, bid Rat antigen-induced arthritis (AIA): Female Lewis rats were sensitized with mBSA and challenged intra-articularly. 13 was administered orally at 30 mg/kg bid for 7 days. Knee swelling was measured with calipers. Mouse angiogenesis chamber model: Female FVB mice were implanted with VEGF-containing chambers and treated with 13 (30 mg/kg bid po) for 4 days. Vascularized tissue weight was measured on day 4. Rat CFA-induced hyperalgesia model: Male Wistar Han rats received intraplantar CFA injection, and after 3 days, 13 was administered orally. Mechanical withdrawal thresholds were measured at 1, 3, and 6 h post-dose. Rat pharmacokinetic study: Female Sprague–Dawley rats received 13 intravenously (1 mg/kg in NMP-PEG200) or orally (3 mg/kg in CMC-Tween-water). Blood samples were collected for concentration analysis.[1] |
| ADME/Pharmacokinetics |
In rats, the clearance (CL) of 13 was 6 ± 1 mL·min⁻¹·kg⁻¹, and the AUC after oral dose standardization was 6.7 ± 2.0 µmol·h·L⁻¹. In the rat AIA model, after administration of 30 mg/kg bid, the plasma concentration at 2 hours was 9.03 ± 2.87 µM, and the plasma concentration at 16 hours was 0.09 ± 0.06 µM. The plasma protein binding rate in rats was 95%, and the plasma protein binding rate in humans was 93%. [1]
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| Toxicity/Toxicokinetics |
13 showed a low hERG binding rate (IC₅₀ = 19 µM) and the smallest inhibition in patch-clamp experiments (44% inhibition at 60 µM).
No significant inhibition was observed in the detection of more than 150 enzymes, receptors and ion channels. Histamine H₃ receptor binding rate >30 µM, indicating a low risk of H₃-related side effects. [1] |
| References |
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| Additional Infomation |
Compound 13 is a selective, orally effective GPR4 antagonist developed from lead compound 1a, with better safety profile in terms of off-target activity against hERG and H₃ receptors. It has shown efficacy in models of inflammation, angiogenesis, and pain, supporting its potential as an anti-inflammatory candidate. Compound 13 is also considered a candidate for treating tissue acidosis-related diseases such as arthritis and chronic pain in certain contexts. [1]
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| Molecular Formula |
C24H28N6O
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|---|---|
| Molecular Weight |
416.518724441528
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| Exact Mass |
416.23
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| Elemental Analysis |
C, 69.21; H, 6.78; N, 20.18; O, 3.84
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| CAS # |
1401728-56-0
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| PubChem CID |
68379135
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| Appearance |
White to yellow solid powder
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| LogP |
3.5
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
31
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| Complexity |
578
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
HXPQWNPLNIEJOW-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C24H28N6O/c1-4-21-20(22-26-15(2)13-16(3)30(22)29-21)14-17-5-7-18(8-6-17)23-27-28-24(31-23)19-9-11-25-12-10-19/h5-8,13,19,25H,4,9-12,14H2,1-3H3
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| Chemical Name |
2-[4-[(2-ethyl-5,7-dimethylpyrazolo[1,5-a]pyrimidin-3-yl)methyl]phenyl]-5-piperidin-4-yl-1,3,4-oxadiazole
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| Synonyms |
NE52QQ57; NE 52-QQ57; NE52-QQ57; NE-52-QQ57; NE-52QQ57; NE 52QQ57
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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 |
| 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: ~20 mg/mL (~48.0 mM)
Ethanol: ~45 mg/mL |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2 mg/mL (4.80 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 20.0 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 mg/mL (4.80 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 20.0 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. View More
Solubility in Formulation 3: ≥ 2 mg/mL (4.80 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.4008 mL | 12.0042 mL | 24.0085 mL | |
| 5 mM | 0.4802 mL | 2.4008 mL | 4.8017 mL | |
| 10 mM | 0.2401 mL | 1.2004 mL | 2.4008 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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