| Size | Price | Stock | Qty |
|---|---|---|---|
| 100mg |
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| 500mg | |||
| Other Sizes |
| Targets |
(Rac)-EC5026 targets soluble epoxide hydrolase (sEH; EPHX2), the enzyme responsible for the hydrolysis of epoxy-fatty acids to their corresponding diols. sEH is a bifunctional enzyme with both epoxide hydrolase and lipid phosphate phosphatase activities, and it is expressed in tissues including liver, kidney, brain, and vascular endothelium. The epoxide hydrolase activity of sEH converts anti-inflammatory epoxy-fatty acids (including EETs, epoxydocosapentaenoic acids, and epoxyeicosatetraenoic acids) to less active diols, thereby terminating their biological activities. By inhibiting sEH, EC5026 preserves the levels of these endogenous lipid mediators, which act as potent vasodilators, anti-inflammatory agents, and analgesics. sEH inhibition represents an attractive therapeutic strategy for pain management because it does not involve opioid receptors and avoids the addiction and respiratory depression risks associated with opioids.
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| ln Vitro |
(Rac)-EC5026 has a Ki of 0.06 nM for inhibiting soluble epoxide hydrolase (sEH)[1].
(Rac)-EC5026 is an exceptionally potent sEH inhibitor with a Ki value of 0.06 nM, representing sub-nanomolar affinity for the enzyme. The compound inhibits sEH activity in a competitive manner, binding to the enzyme's active site and preventing the hydrolysis of epoxy-fatty acids. In cell-based assays, EC5026 at picomolar to low nanomolar concentrations effectively increases intracellular levels of EETs and reduces the production of pro-inflammatory cytokines (TNF-α, IL-6, IL-1β) and chemokines in activated immune cells. The compound shows excellent selectivity for sEH over other epoxide hydrolases and a panel of >50 off-target enzymes, receptors, and ion channels. The racemic mixture demonstrates potent anti-inflammatory activity in vitro, comparable to the single enantiomer forms, indicating that both enantiomers contribute to the overall efficacy. |
| ln Vivo |
In preclinical models, (Rac)-EC5026 has demonstrated robust efficacy as a non-opioid analgesic. In the formalin-induced paw licking model of inflammatory pain, oral administration of EC5026 at 0.3-10 mg/kg significantly reduces both the early and late phases of pain behavior. In the chronic constriction injury (CCI) model of neuropathic pain, the compound (1-10 mg/kg, p.o.) attenuates mechanical allodynia and thermal hyperalgesia. The analgesic efficacy of EC5026 is comparable to that of gabapentin and pregabalin, but with a different mechanism of action. In the spinal nerve ligation (SNL) model, EC5026 reduces pain behaviors and normalizes spinal cord levels of inflammatory mediators. The compound also shows efficacy in models of osteoarthritis and chemotherapy-induced neuropathic pain. Importantly, EC5026 does not produce tolerance, dependence, or respiratory depression, unlike opioid analgesics.
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| Enzyme Assay |
sEH inhibitory activity of (Rac)-EC5026 is measured using a fluorescence-based enzyme assay with recombinant human sEH. The assay utilizes a fluorescent substrate (PHOME) that releases a fluorescent product upon hydrolysis by sEH. The enzyme (0.1-0.5 nM) is incubated with varying concentrations of the test compound (0.001-100 nM) and substrate (5-10 μM) in assay buffer (25 mM Tris-HCl, pH 7.4, 0.1 mg/mL BSA) at 30°C for 15-30 minutes. The reaction is quenched with acidic stop solution, and fluorescence is measured at excitation/emission 330/465 nm. The IC50 is calculated from dose-response curves, and the Ki is determined using the Cheng-Prusoff equation. The binding affinity is also assessed by surface plasmon resonance (SPR) with immobilized sEH. Selectivity is evaluated by screening against a panel of 50-100 related enzymes, receptors, and ion channels.
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| Cell Assay |
The cellular activity of (Rac)-EC5026 is evaluated in human and rodent cell lines. Primary macrophages, microglia, or dorsal root ganglion (DRG) neurons are treated with the compound at concentrations ranging from 0.01 to 100 nM for 1-24 hours. After treatment, cells are stimulated with inflammatory mediators (LPS, IL-1β, or TNF-α) or pain-inducing stimuli (capsaicin, bradykinin). EET and DHET levels are quantified by LC-MS/MS. Cytokine and chemokine levels are measured by ELISA or multiplex bead-based assays. Neuronal excitability is assessed by patch-clamp electrophysiology or calcium imaging. The compound's effect on inflammatory signaling pathways (NF-κB, MAPK, PI3K/AKT) is evaluated by Western blot analysis of phosphorylated signaling proteins. Cell viability and cytotoxicity are assessed by MTT and LDH release assays.
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| Animal Protocol |
In animal efficacy and pharmacokinetic studies, (Rac)-EC5026 is administered to rodents (mice or rats) via oral gavage, intraperitoneal injection, or subcutaneous injection at doses of 0.3-30 mg/kg. For pain models, drug administration is typically 30-120 minutes before pain assessment, and pain behaviors are measured using von Frey filaments, hot plate, tail flick, or formalin tests. For chronic pain models, animals are treated daily for 7-28 days, with pain assessments conducted at regular intervals. Blood samples are collected at various time points for pharmacokinetic analysis. At study termination, tissues (liver, kidney, brain, spinal cord, DRG) are harvested for measurement of EET/DHET levels by LC-MS/MS, inflammatory markers by ELISA, and histopathological examination. The compound's effect on opioid receptors is assessed by naloxone challenge experiments to confirm the non-opioid mechanism of action.
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| ADME/Pharmacokinetics |
Preclinical pharmacokinetic studies of (Rac)-EC5026 in rodents show that the compound is orally bioavailable with excellent absorption and favorable half-life. Following oral administration at 1-10 mg/kg, the compound achieves peak plasma concentrations (Cmax) within 1-2 hours (Tmax) and has a plasma half-life of 4-8 hours in rats and 2-4 hours in mice. The oral bioavailability is approximately 60-80%, supporting once- or twice-daily dosing. The compound shows low plasma protein binding (<50%) and a moderate volume of distribution (Vd ~1-2 L/kg), indicating good tissue penetration. The compound crosses the blood-brain barrier, achieving brain concentrations sufficient to modulate central sEH activity, which is important for its analgesic effects. Metabolism is primarily via CYP450 enzymes (CYP3A4, CYP2C9), and the compound is excreted in urine and feces. The racemic mixture exhibits similar PK parameters to the single enantiomers, suggesting that the enantiomers do not significantly interconvert in vivo.
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| Toxicity/Toxicokinetics |
Toxicology studies of (Rac)-EC5026 have been conducted in rodents and non-human primates as part of its preclinical development. At therapeutic doses (0.3-10 mg/kg/day), the compound is well-tolerated in 14-day and 28-day repeat-dose studies. No significant adverse effects on body weight, food consumption, hematology, clinical chemistry, or histopathology have been observed. At high doses (>30 mg/kg/day), mild to moderate increases in liver enzymes (ALT, AST) have been reported, along with hepatocellular hypertrophy, likely due to metabolic enzyme induction rather than direct hepatotoxicity. No genotoxicity or carcinogenicity has been detected in standard assays. The compound does not affect the central nervous system at therapeutic doses, as assessed by functional observation battery (FOB) tests. The no-observed-adverse-effect level (NOAEL) is established at 10-30 mg/kg/day, providing a safety margin of approximately 10-30 fold for clinical studies. EC5026 is being developed as a non-opioid analgesic, and its safety profile is considered favorable for chronic pain management.
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| References | |
| Additional Infomation |
(Rac)-EC5026 (BPN-19186) is a first-in-class, non-opioid sEH inhibitor being developed for the treatment of inflammatory and neuropathic pain. The compound's mechanism of action—stabilizing endogenous analgesic epoxy-fatty acids—represents a novel approach to pain management that avoids the addiction, tolerance, and respiratory depression risks associated with opioid drugs. (Rac)-EC5026 has advanced into clinical development, with Phase 1 and Phase 2 trials conducted to evaluate its safety, tolerability, and efficacy in patients with pain conditions. The compound is also being investigated for potential applications in Parkinson's disease and dementia with Lewy bodies (DLB), based on the role of epoxy-fatty acids in neuroinflammation and neurodegeneration. The development of EC5026 highlights the therapeutic potential of targeting sEH for pain management and provides a promising alternative to opioids in an era of increasing concern about opioid addiction and overdose.
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| Molecular Formula |
C18H23F4N3O3
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|---|---|
| Molecular Weight |
405.387138605118
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| Exact Mass |
405.167
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| CAS # |
1809885-55-9
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| Related CAS # |
EC5026;1809885-32-2
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| PubChem CID |
118395966
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| Appearance |
White to off-white solid powder
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| LogP |
3.7
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
7
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
28
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| Complexity |
539
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| Defined Atom Stereocenter Count |
0
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| SMILES |
N(C1=CC=C(OC(F)(F)F)C(F)=C1)C(NC1CCN(C(=O)C(C)CC)CC1)=O
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| InChi Key |
LHRXHTKENPCGSZ-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C18H23F4N3O3/c1-3-11(2)16(26)25-8-6-12(7-9-25)23-17(27)24-13-4-5-15(14(19)10-13)28-18(20,21)22/h4-5,10-12H,3,6-9H2,1-2H3,(H2,23,24,27)
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| Chemical Name |
1-[3-fluoro-4-(trifluoromethoxy)phenyl]-3-[1-(2-methylbutanoyl)piperidin-4-yl]urea
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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 : ~250 mg/mL (~616.69 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (5.13 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 20.8 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: ≥ 2.08 mg/mL (5.13 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 20.8 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: ≥ 2.08 mg/mL (5.13 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 | 2.4668 mL | 12.3338 mL | 24.6676 mL | |
| 5 mM | 0.4934 mL | 2.4668 mL | 4.9335 mL | |
| 10 mM | 0.2467 mL | 1.2334 mL | 2.4668 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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