| Size | Price | Stock | Qty |
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| 50mg |
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| Other Sizes |
| Targets |
The primary molecular target of 3-AQC is the 5-HT3 receptor (also known as the 5-HT3A channel), a ligand-gated ion channel of the Cys-loop family. This receptor is a pentameric structure with five subunits surrounding a central ion channel pore. Upon binding of serotonin (5-HT), the channel opens, allowing cation influx and neuronal depolarization. 3-AQC functions as a competitive antagonist at this receptor, meaning it binds reversibly to the same orthosteric site as serotonin, thereby blocking serotonin’s action without activating the receptor itself. This competitive mechanism underlies its utility as a pharmacological tool for studying serotonergic signaling pathways involved in emesis, anxiety, and gastrointestinal motility.
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| ln Vitro |
3-AQC (Compound 7e) has about 100 times the potency of tropisetron. In the ileum of guinea pigs, the pA2 for 2-methyl-5HT antagonistic activity is 10.2 [1].
In vitro, 3-AQC demonstrates exceptionally high antagonist potency at the 5-HT3 receptor. The compound (referred to as compound 7e in the original synthesis publication) exhibits approximately 100-fold greater potency compared to the standard 5-HT3 antagonist tropisetron. In functional assays using isolated guinea pig ileum, 3-AQC potently antagonizes contractions induced by the selective 5-HT3 agonist 2-methyl-5-HT, with a pA2 value of 10.2, indicating a very high affinity and potency at the receptor in this native tissue preparation. The compound is reported to be highly selective for the 5-HT3A receptor subtype, with continued binding assays confirming its competitive mode of antagonism. |
| ln Vivo |
While detailed in vivo pharmacokinetic datasets for 3-AQC are limited in publicly available literature, functional tissue-based assays are also considered to reflect ex vivo activity relevant to in vivo applications. Notably, 3-AQC has been reported to exhibit widely differing antagonist activity across various tissues, suggesting potential tissue-specific factors influencing its in vivo efficacy beyond simple receptor affinity. This tissue-dependent activity profile underscores the importance of careful experimental design when using 3-AQC in vivo. The compound has been utilized as a pharmacological tool in developmental biology studies, including investigations of the pre-nervous serotonergic system in developing sea urchin embryos and larvae, indicating its applicability in whole-organism research settings beyond standard mammalian models.
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| Enzyme Assay |
The classical receptor binding and functional assay protocol for 5-HT3 antagonists like 3-AQC utilizes the isolated guinea pig ileum preparation. Briefly, segments of guinea pig ileum (approximately 2–3 cm) are mounted in organ baths containing oxygenated physiological salt solution maintained at 37°C. Tissues are equilibrated under 1 g tension for 45–60 minutes with regular washing. Cumulative concentration-response curves to the selective 5-HT3 agonist 2-methyl-5-HT are constructed. 3-AQC is then added to the bath at various concentrations and allowed to incubate for a set period (typically 20–30 minutes) before re-determining the agonist concentration-response curve in the presence of the antagonist. Antagonist potency is quantified by calculating the pA2 value (negative logarithm of the antagonist concentration that produces a 2-fold shift in the agonist EC50) using Schild regression analysis. For electrophysiological characterization of 5-HT3A channel blockade, whole-cell patch-clamp recordings can be performed on heterologously expressing cell lines, where currents are elicited by 10 µM 5-HT delivered every 3 minutes, and 3-AQC is applied 30 seconds before stimulation at concentrations ranging from 10 to 100 µM to demonstrate dose-dependent and reversible inhibition.
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| Cell Assay |
Cellular assays for 5-HT3 antagonists are typically performed using cell lines heterologously expressing the human 5-HT3A receptor (e.g., HEK293 or CHO cells). Standard procedure: Cells are cultured in appropriate medium (e.g., DMEM with 10% FBS and selection antibiotics) at 37°C in a 5% CO₂ humidified incubator. For functional calcium influx assays, cells are seeded in 96-well black-wall clear-bottom plates at approximately 3–5 × 10⁴ cells per well and allowed to adhere overnight. The next day, medium is removed, and cells are loaded with a calcium-sensitive fluorescent dye (e.g., Fluo-4 AM) in assay buffer (Hanks‘ Balanced Salt Solution with 20 mM HEPES, pH 7.4) for 45–60 minutes at 37°C. After washing, varying concentrations of 3-AQC are added for 10–15 minutes pre-incubation. Fluorescence signal is then recorded upon addition of EC₈₀ concentration of 5-HT or the selective agonist 2-methyl-5-HT using a fluorescence plate reader. Alternatively, whole-cell patch-clamp electrophysiology can be employed: cells are continuously perfused with extracellular solution, 5-HT (10 µM) is applied every 3 minutes to elicit currents, and 3-AQC is applied 30 seconds before each 5-HT stimulation at concentrations ranging from 10 to 100 µM to quantify inhibition in a dose-dependent manner.
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| Animal Protocol |
Detailed animal in vivo protocols for 3-AQC are not extensively reported in public literature. However, general in vivo experimental design considerations for 5-HT3 antagonists can be adapted. Typically, rodent models (mice or rats) are used. For pharmacokinetic studies, animals are administered 3-AQC via intravenous (IV) or oral (PO) routes; blood samples are collected at predetermined time points post-administration (e.g., 0, 5, 15, 30 min; 1, 2, 4, 8, 12, 24 h) from the tail vein or via cardiac puncture at terminal time points. Plasma concentrations are quantified using validated LC-MS/MS methods. Pharmacodynamic studies may employ behavioral paradigms sensitive to serotonergic modulation, such as the elevated plus maze for anxiety-like behavior or conditioned taste aversion for emesis models. The compound is typically dissolved in a suitable vehicle — for example, a formulation of 10% DMSO + 40% PEG300 + 5% Tween 80 + 45% saline — and administered via intraperitoneal (IP), intravenous, or oral routes. Dose conversion between species follows the FDA-recommended Km factor method (e.g., mouse Km=3, rat Km=6).
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| ADME/Pharmacokinetics |
Systematic and comprehensive pharmacokinetic data (e.g., oral bioavailability, half-life, volume of distribution, clearance) for 3-AQC in animal models or humans are not readily available in the currently accessible literature. The compound‘s solubility is reported as soluble to 100 mM in DMSO and up to 11 mg/mL (27.82 mM) in DMSO under sonication, with aqueous solubility likely limited given its lipophilic heterocyclic structure. The compound is typically stored as a powder at -20°C, where it remains stable for up to 3 years; working solutions in solvent can be stored at -80°C for 1 year.
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| Toxicity/Toxicokinetics |
Available Safety Data Sheets (SDS) indicate that 3-AQC is not classified as a hazardous substance or mixture under standard criteria; acute toxicity data are listed as not available. However, it is noted that under fire conditions, the compound may decompose and emit toxic fumes, and users should avoid release into the environment. The compound is intended strictly for laboratory research purposes and is not approved for human use. Standard safety precautions should be observed during handling, including appropriate personal protective equipment (lab coat, gloves, safety goggles) and working within a properly ventilated fume hood. Note that there exists a distinct compound also abbreviated as “3AQC” (3-aminoquinoxaline-2-carbonitrile, CAS 6635-31-0), which is reported to exhibit cytotoxicity in vitro against V79 cells and other cell lines, and may cause DNA damage, as well as demonstrate bioreductive effects on solid tumors. This cytotoxic compound is chemically distinct from the 5-HT3 antagonist 3-AQC described throughout this report.
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| References |
| Molecular Formula |
C20H21N5O4
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|---|---|
| Molecular Weight |
395.41
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| Exact Mass |
395.159
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| CAS # |
201216-42-4
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| PubChem CID |
11567402
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| Appearance |
Typically exists as solid at room temperature
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| LogP |
1.524
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
9
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
29
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| Complexity |
525
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C=CCN1CCN(C2=NC3=CC=CC=C3N=C2C#N)CC1.OC(/C=C\C(=O)O)=O
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| InChi Key |
UHLVYEOCPBNJNA-BTJKTKAUSA-N
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| InChi Code |
InChI=1S/C16H17N5.C4H4O4/c1-2-7-20-8-10-21(11-9-20)16-15(12-17)18-13-5-3-4-6-14(13)19-16;5-3(6)1-2-4(7)8/h2-6H,1,7-11H2;1-2H,(H,5,6)(H,7,8)/b;2-1-
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| Chemical Name |
3-(4-Prop-2-enylpiperazin-1-yl)quinoxaline-2-carbonitrile Maleate
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| Synonyms |
3 AQC 3AQC 3-AQC
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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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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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 | 2.5290 mL | 12.6451 mL | 25.2902 mL | |
| 5 mM | 0.5058 mL | 2.5290 mL | 5.0580 mL | |
| 10 mM | 0.2529 mL | 1.2645 mL | 2.5290 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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