| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
5-HT1A Receptor 34 nM (IC50)
5-HT1A receptor (antagonist, IC50 values: 15 nM (potent), 34 nM at rat hippocampal 5-HT1A receptor) |
|---|---|
| ln Vitro |
With a pA2 of 7.2, WAY100135 (0.1-1 μM) inhibits 5-carboxamidoiodotryptamine electrically induced contractions in the ileum of guinea pigs[1]. The inhibitory effect of 8-OH-DPAT on eEPSC and eIPSC in MVM neurons is blocked by WAY-100135 dihydrochloride (10 μM)[2].
In radioligand binding assays, WAY-100135 dihydrochloride is a potent, selective 5-HT1A receptor antagonist with an IC50 of 15 nM. It is highly selective over 5-HT1B, 5-HT1C, 5-HT2, alpha1, alpha2, and D2 receptors, with IC50 values greater than 1000 nM for these off-target sites, demonstrating its excellent selectivity for the 5-HT1A receptor. |
| ln Vivo |
In rats under anesthesia, WAY100135 (2.5 mg/kg) inhibits raphe neuronal activity up to a maximum of 30% and counteracts the firing suppression caused by 8-OH-DPAT (0.5 mg/kg iv)[1]. Rats' behavioral deficits generated by 5-HT are successfully eliminated by WAY-100135 dihydrochloride (10 mM; 1 μL for microinjection)[2].
WAY-100135 dihydrochloride is a selective antagonist at both presynaptic and postsynaptic 5-HT1A receptors. It has been used in numerous in vivo studies to investigate the role of 5-HT1A autoreceptors in the regulation of serotonin release and the therapeutic effects of SSRIs and other serotonergic antidepressants. Blockade of presynaptic 5-HT1A autoreceptors enhances serotonin release in the forebrain. |
| Enzyme Assay |
Standard radioligand binding assays for 5-HT1A receptors are performed using membrane preparations from rat hippocampus (which abundantly expresses 5-HT1A receptors) or from CHO cells expressing human 5-HT1A receptors. [3H]-8-OH-DPAT (a selective 5-HT1A agonist) is used as the radioligand (final concentration ~0.5-1 nM). Test compound (WAY-100135) is incubated with membranes at varying concentrations (0.1 nM to 10 uM) for 60 min at 25degC. Nonspecific binding is determined using 10 uM serotonin or 10 uM 8-OH-DPAT. Bound radioligand is separated by vacuum filtration through glass fiber filters and counted by liquid scintillation. IC50 values are calculated from competition curves. Ki values are then derived using the Cheng-Prusoff equation.
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| Cell Assay |
Cell-based functional assays for 5-HT1A antagonism are typically performed using CHO (Chinese hamster ovary) cells stably expressing human 5-HT1A receptors. The 5-HT1A receptor is Gi/o-coupled; its activation inhibits forskolin-stimulated cAMP accumulation. For antagonist assays, cells are seeded in 96-well plates. The next day, cells are treated with serial dilutions of WAY-100135 dihydrochloride (1 nM to 10 uM) in the presence of forskolin (typically 1-10 uM, which directly activates adenylyl cyclase). A selective 5-HT1A agonist (e.g., 8-OH-DPAT at its EC80 concentration, typically 10-100 nM) is added to activate the receptor and inhibit cAMP production. After 15-30 min, intracellular cAMP levels are quantified using a HTRF (Homogeneous Time-Resolved Fluorescence) or ELISA-based cAMP detection kit. Antagonist potency (IC50) is determined as the concentration that reverses 50% of the agonist-mediated inhibition of cAMP accumulation. IC50 values for WAY-100135 are typically 15-34 nM. EC50 for agonist activity is not applicable (pEC50 = not applicable).
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| Animal Protocol |
In vivo animal studies are typically conducted in rodents (male Sprague-Dawley rats or C57BL/6 mice, 8-12 weeks old). WAY-100135 dihydrochloride is dissolved in sterile saline or water and administered systemically via intraperitoneal (i.p.) injection at doses ranging from 0.1 to 10 mg/kg, or locally by intracerebral microinjection (nmol range). For behavioral studies, the compound is administered 15-60 min prior to testing. Experimental models include: (1) Forced swim test and tail suspension test for antidepressant-like effects; (2) Elevated plus maze and open field test for anxiety; (3) Microdialysis to measure extracellular serotonin release in the forebrain (e.g., medial prefrontal cortex or ventral hippocampus). Pharmacodynamic parameters include changes in immobility time (seconds), percentage of open arm entries, and extracellular 5-HT levels (expressed as percentage of baseline). The antagonist effect is typically confirmed by reversing the effects of a 5-HT1A agonist such as 8-OH-DPAT (e.g., reversal of hypothermia, behavioral effects, or inhibition of serotonin release).
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| ADME/Pharmacokinetics |
Dosing and exposure: Following i.p. administration in rats at a typical research dose of 1-3 mg/kg, WAY-100135 shows adequate brain penetration to occupy central 5-HT1A receptors. Pharmacokinetic parameters (e.g., plasma half-life t1/2, Cmax, AUC, clearance CL, volume of distribution Vd) for this specific research compound are not fully standardized. However, as a small molecule that is not a clinical drug, PK studies for this antagonist are generally not required; in vivo efficacy is empirically determined from dose-response and time-course experiments.
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| Toxicity/Toxicokinetics |
The compound is a research tool and not intended for human therapeutic use; therefore, detailed regulatory toxicology studies are not publicly available. In acute rodent studies, typical research doses (1-10 mg/kg i.p.) are well tolerated without overt signs of toxicity. As a specific and selective 5-HT1A antagonist, it does not exhibit the significant cardiotoxicity or cytotoxicity often associated with non-specific drugs. However, high doses may produce behavioral effects due to its mechanism of action. Standard safety precautions include using appropriate personal protective equipment (PPE) when handling the powder.
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| References |
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| Additional Infomation |
N-tert-butyl-3-[4-(2-methoxyphenyl)-1-piperazinyl]-2-phenylpropionamide is a member of the piperazine class of compounds.
WAY-100135 dihydrochloride is a seminal tool compound for 5-HT1A receptor research. It was one of the first highly selective, silent antagonists (i.e., possessing no intrinsic activity itself) available for this receptor. Its selectivity over 5-HT1B, 5-HT1C, 5-HT2, alpha1, alpha2 and D2 receptors (>1000 nM) makes it invaluable for dissecting 5-HT1A-mediated functions. The compound is not a drug and is for research use only. It is used primarily to differentiate between presynaptic (autoreceptor) and postsynaptic 5-HT1A receptor functions, and to investigate the role of 5-HT1A receptors in the mechanism of action of SSRIs. Key synonyms include WAY100135 dihydrochloride and WAY 100135. The pharmacological profile is well-established, with an IC50 of 15 nM (most potent) or 34 nM (at rat hippocampal 5-HT1A receptor) depending on the specific assay conditions. |
| Molecular Formula |
C24H35CL2N3O2
|
|---|---|
| Molecular Weight |
468.46
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| Exact Mass |
467.211
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| CAS # |
149055-79-8
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| PubChem CID |
115111
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| Appearance |
Typically exists as solid at room temperature
|
| LogP |
5.962
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
7
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| Heavy Atom Count |
29
|
| Complexity |
505
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
Cl.Cl.COC1=CC=CC=C1N1CCN(CC(C2=CC=CC=C2)C(NC(C)(C)C)=O)CC1
|
| InChi Key |
UMTDAKAAYOXIKU-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C24H33N3O2/c1-24(2,3)25-23(28)20(19-10-6-5-7-11-19)18-26-14-16-27(17-15-26)21-12-8-9-13-22(21)29-4/h5-13,20H,14-18H2,1-4H3,(H,25,28)
|
| Chemical Name |
N-tert-butyl-3-[4-(2-methoxyphenyl)piperazin-1-yl]-2-phenylpropanamide
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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) |
H2O: 5 mg/mL (10.67 mM)
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|---|---|
| Solubility (In Vivo) |
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.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *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). View More
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)] Oral Formulations
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). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.1347 mL | 10.6733 mL | 21.3465 mL | |
| 5 mM | 0.4269 mL | 2.1347 mL | 4.2693 mL | |
| 10 mM | 0.2135 mL | 1.0673 mL | 2.1347 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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