| Size | Price | Stock | Qty |
|---|---|---|---|
| 10mg |
|
||
| 50mg |
|
||
| Other Sizes |
| Targets |
Trace amine-associated receptor 1 (TAAR1). RO 5212773 (EPPTB) acts as a selective antagonist for mouse TAAR1, with a reported Ki of 0.9 nM and an IC50 of 28 nM. It has no significant effects on other TAAR subtypes, making it a highly specific tool for studying TAAR1-mediated pathways.
|
|---|---|
| ln Vitro |
Hippocampal pyramidal neurons' excitability is inhibited by Ro 5212773 (EPPTB). Additionally, EPPTB lowers seizure activity and epileptiform events (SLE) [1]. Inward rectifier K+ current activation mediated by TAAR1 is blocked by Ro 5212773 [2]. 1.5 μM β-phenylethylamine (IC50=27.5 nM) potently inhibits the production of cAMP induced by mouse TAAR1 activation. This is achieved by Ro 5212773. In the absence of TAAR1 agonist, Ro 5212773 dose-dependently lowers cAMP levels in HEK293 cells (IC50= 19 nM). Compared to rat (IC50= 4539 nM) and human (IC50= 7487 nM) TAAR1, EPPTB significantly more effectively inhibits the production of cAMP in mice [2].
In a cell-based assay using HEK293 cells expressing mouse TAAR1, EPPTB potently antagonizes cAMP production induced by the non-selective TAAR1 agonist beta-phenylethylamine with an IC50 of 27.5 nM. It also blocks the reduction of dopamine (DA) neuron firing frequency induced by the TAAR1 agonist p-tyramine, confirming its functional antagonist activity. |
| ln Vivo |
In vivo, EPPTB effectively prevents the reduction in the firing frequency of dopamine (DA) neurons in the ventral tegmental area (VTA) induced by the TAAR1 agonist p-tyramine. This demonstrates its ability to cross the blood-brain barrier and act on central TAAR1 targets. It may be a promising target for studying neurodegenerative disorders.
|
| Enzyme Assay |
A standard competitive binding assay is performed. Membranes from cells expressing mouse TAAR1 are incubated with a high-affinity radiolabeled ligand (e.g., [3H]RO5212773) and varying concentrations of unlabeled EPPTB. Bound and free ligands are separated by filtration, and the Ki value is calculated.
|
| Cell Assay |
A functional cAMP assay is performed. HEK293 cells stably expressing mouse TAAR1 are seeded in 96-well plates. Cells are pre-incubated with EPPTB, then stimulated with a selective TAAR1 agonist. After cell lysis, intracellular cAMP levels are quantified using a HTRF or chemiluminescence-based immunoassay to determine the antagonist's IC50.
|
| Animal Protocol |
For electrophysiological studies, male C57BL/6J mice are used. EPPTB is dissolved in a suitable vehicle and administered via intraperitoneal (i.p.) or intracerebroventricular (i.c.v.) injection. In vivo extracellular recordings of dopamine neuron firing in the ventral tegmental area (VTA) are performed to assess the compound's ability to reverse the effects of a TAAR1 agonist.
|
| ADME/Pharmacokinetics |
EPPTB is described as having good bioavailability in vivo and the ability to cross the blood-brain barrier, as demonstrated by its central effects on dopamine neuron firing. However, no specific numerical PK parameters (e.g., t1/2, Cmax) have been reported in the available literature.
|
| Toxicity/Toxicokinetics |
Not reported. As a research tool, no formal toxicology data is available. However, it is considered a "useful tool for probing the physiologic role of TAAR1" and its effects are well-tolerated in standard animal study protocols.
|
| References | |
| Additional Infomation |
RO 5212773 (EPPTB) is a pivotal compound in TAAR1 research. TAAR1 is a G protein-coupled receptor (GPCR) that is activated by various endogenous trace amines and is a promising drug target for psychiatric disorders such as schizophrenia and addiction. EPPTB's discovery allowed researchers to delineate TAAR1-specific functions from other monoamine systems, revealing its role as a key regulator of dopamine and glutamate neurotransmission.
|
| Molecular Formula |
C20H21F3N2O2
|
|---|---|
| Molecular Weight |
378.39
|
| Exact Mass |
378.156
|
| CAS # |
1110781-88-8
|
| PubChem CID |
25175634
|
| Appearance |
White to off-white solid powder
|
| LogP |
5.094
|
| Hydrogen Bond Donor Count |
1
|
| Hydrogen Bond Acceptor Count |
6
|
| Rotatable Bond Count |
5
|
| Heavy Atom Count |
27
|
| Complexity |
494
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
CCOC1=CC=CC(=C1)NC(=O)C2=CC(=C(C=C2)N3CCCC3)C(F)(F)F
|
| InChi Key |
KLFVWQCQUXXLOU-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C20H21F3N2O2/c1-2-27-16-7-5-6-15(13-16)24-19(26)14-8-9-18(25-10-3-4-11-25)17(12-14)20(21,22)23/h5-9,12-13H,2-4,10-11H2,1H3,(H,24,26)
|
| Chemical Name |
N-(3-ethoxyphenyl)-4-pyrrolidin-1-yl-3-(trifluoromethyl)benzamide
|
| HS Tariff Code |
2934.99.9001
|
| 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)
|
| Solubility (In Vitro) |
DMSO: 100 mg/mL (264.28 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (6.61 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.6428 mL | 13.2139 mL | 26.4278 mL | |
| 5 mM | 0.5286 mL | 2.6428 mL | 5.2856 mL | |
| 10 mM | 0.2643 mL | 1.3214 mL | 2.6428 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
