| Size | Price | Stock | Qty |
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| 50mg |
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| 100mg |
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| 250mg |
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| Other Sizes |
| Targets |
- γ-Aminobutyric acid (GABA) receptors or GABAergic system [1]
- Human proton-coupled amino acid transporter 1 (hPAT1) (served as a substrate of hPAT1; pH-dependent transport activity was confirmed) [2] |
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| ln Vitro |
- Experiments were performed on synaptosomal preparations or brain slices isolated from cats to investigate the effect of Arecaidine on GABA-mediated inhibitory responses in the central nervous system. When Arecaidine was applied together with GABA, it was observed to antagonize the inhibitory postsynaptic potentials (IPSPs) induced by GABA. Specifically, Arecaidine reduced both the amplitude and duration of GABA-evoked IPSPs in neurons from cat central nervous system regions (e.g., cerebral cortex and thalamus). This result suggested that Arecaidine could interfere with GABAergic inhibitory neurotransmission. [1]
- In HEK293 cells transfected with the human hPAT1 gene (empty vector-transfected cells as control), the transport of Arecaidine was evaluated using radiolabeled tracers. The results showed that Arecaidine was actively transported by hPAT1, and this transport exhibited pH dependence—significantly higher uptake of Arecaidine was detected under acidic conditions (mimicking the environment of the intestinal lumen) compared to neutral conditions. Additionally, the uptake of Arecaidine by hPAT1-transfected cells was competitively inhibited by excess unlabeled hPAT1 substrates (such as GABA and L-proline), which confirmed that the transport of Arecaidine was mediated specifically by hPAT1. [2] |
| ln Vivo |
- Adult cats were used as experimental animals. After anesthesia, intracranial microelectrodes were implanted into specific brain regions (e.g., cerebral cortex and thalamus) of the cats to record extracellular electrical activities of neurons. Arecaidine was dissolved in artificial cerebrospinal fluid and administered via intracerebral microinjection near the recorded neurons. After administration of Arecaidine, the inhibitory effect of exogenously applied GABA on neuronal firing was weakened—manifested as an increase in the discharge frequency of neurons compared to the group treated with GABA alone. The antagonistic effect of Arecaidine lasted for approximately 15-30 minutes, and the neuronal electrical activity gradually recovered thereafter. [1]
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| Enzyme Assay |
- GABA receptor binding assay: Brain homogenates were prepared from the cerebral cortex of cats, serving as the source of GABA receptors. Radiolabeled [³H]-GABA (as a ligand) was mixed with the brain homogenates and different concentrations of Arecaidine in binding buffer. The mixture was incubated at 4°C for a certain period, then filtered through glass fiber filters to separate the bound [³H]-GABA from the free form. The radioactivity of the filters (representing bound [³H]-GABA) was measured using a liquid scintillation counter. The results indicated that Arecaidine could compete with [³H]-GABA for binding to GABA receptors, as it reduced the specific binding of [³H]-GABA in a concentration-dependent manner, confirming the interaction between Arecaidine and GABA receptors. [1]
- hPAT1-mediated transport assay: HEK293 cells were transfected with hPAT1 expression plasmids (or empty vectors as controls) and cultured for 48 hours. The cells were harvested and resuspended in HBSS buffer adjusted to different pH values (acidic and neutral). Radiolabeled Arecaidine was added to the cell suspension, and the mixture was incubated at 37°C for different time intervals. The reaction was terminated by adding ice-cold HBSS buffer, followed by centrifugation to collect the cells. The cells were lysed, and the radioactivity in the cell lysate was measured. The protein concentration of the lysate was determined to normalize the uptake amount of Arecaidine. For competition experiments, excess unlabeled hPAT1 substrates (e.g., GABA, L-proline) were co-incubated with radiolabeled Arecaidine to verify the specificity of hPAT1-mediated transport. [2] |
| Cell Assay |
- Primary cat neuron culture assay: Neurons were isolated from the cerebral cortex of neonatal cats and cultured in appropriate neurobasal medium for 7-10 days. The effect of Arecaidine on GABA-induced calcium responses was detected using the calcium-sensitive dye Fura-2 AM. The neurons were loaded with Fura-2 AM for 30 minutes, then treated with GABA alone or GABA combined with Arecaidine. The fluorescence intensity of Fura-2 AM (excitation at 340 nm and 380 nm, emission at 510 nm) was recorded using a fluorescence microscope. It was found that Arecaidine decreased the GABA-induced calcium influx in neurons, which was consistent with its GABA receptor antagonistic effect. [1]
- hPAT1-expressing HEK293 cell uptake assay: HEK293 cells transfected with hPAT1 were seeded into 24-well plates and cultured until reaching 80% confluence. Before the experiment, the cells were washed twice with HBSS buffer of different pH values. Radiolabeled Arecaidine in HBSS buffer was added to each well, and the cells were incubated at 37°C for a specific time. The incubation was stopped by aspirating the medium and washing the cells with ice-cold HBSS buffer. The cells were lysed with NaOH, and the radioactivity in the lysate was counted. The results showed that the uptake of radiolabeled Arecaidine by hPAT1-transfected cells was significantly higher than that by empty vector-transfected cells, and this uptake was markedly inhibited by co-incubation with unlabeled GABA (a known hPAT1 substrate), confirming the role of hPAT1 in Arecaidine transport. [2] |
| Animal Protocol |
- Adult cats (weight not specified) were anesthetized with pentobarbital sodium (induction via intraperitoneal injection, maintenance via continuous intravenous infusion). The anesthetized cats were fixed in a stereotaxic frame, and a craniotomy was performed to expose the target brain regions (cerebral cortex or thalamus). Glass microelectrodes filled with 3 M NaCl were inserted into the brain tissue to record extracellular potentials of single neurons. Arecaidine was dissolved in artificial cerebrospinal fluid (composition: 126 mM NaCl, 2.5 mM KCl, 1.2 mM MgCl₂, 1.2 mM CaCl₂, 1.0 mM NaH₂PO₄, 26 mM NaHCO₃, 10 mM glucose) to prepare solutions of different concentrations. Microinjections of Arecaidine (volume: 0.5-1 μL) were delivered to the area near the recorded neuron using a microsyringe. The electrical activity of the neuron was recorded continuously for 60 minutes (15 minutes before injection and 45 minutes after injection) to monitor the effect of Arecaidine on GABA-induced inhibition. After the experiment, the cats were euthanized with an overdose of pentobarbital sodium. [1]
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| ADME/Pharmacokinetics |
Arecoline was identified as a substrate of hPAT1, which is highly expressed on the apical membrane of intestinal epithelial cells. This finding suggests that arecoline may be absorbed from the gastrointestinal tract via hPAT1-mediated transport, particularly in the acidic environment of the stomach and upper small intestine (where the proton gradient drives hPAT1 activity). However, in vivo ADME parameters of arecoline (e.g., absorption rate, bioavailability) have not been determined. [2]
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| References |
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| Additional Infomation |
Arecaidine is a naturally occurring pyridine alkaloid isolated from the fruit of Areca catechu L., a common component of areca nut chews. This study focuses on the effects of arecaidine on the GABAergic system of the central nervous system—GABA is the main inhibitory neurotransmitter of the mammalian central nervous system. The results showed that arecaidine, as a GABA receptor antagonist, may enhance the excitatory effects of areca nut (e.g., increased alertness and euphoria) by reducing GABA-mediated central inhibition. [1] Human hPAT1 (SLC36A1) belongs to the solute carrier 36 family, which mediates the uptake of small molecule neutral amino acids, imino acids and certain neurotransmitters (e.g., GABA) via proton-coupled electrogenesis. In addition to intestinal absorption, hPAT1 is also expressed in the proximal tubules of the kidney and may be involved in the reabsorption of filtered arecaidine. Arecoline has been identified as a substrate of hPAT1, which helps to understand the molecular mechanisms of its absorption and distribution in the human body. It also suggests that there may be drug interactions if arecoline is used in combination with other hPAT1 substrates (such as certain amino acid supplements and GABA analogs). [2]
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| Molecular Formula |
C7H11NO2.HCL
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|---|---|
| Molecular Weight |
177.62868
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| Exact Mass |
177.056
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| CAS # |
6018-28-6
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| Related CAS # |
Arecaidine;499-04-7;Arecaidine hydrobromide;6013-57-6
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| PubChem CID |
12305194
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| Appearance |
White to light yellow solid powder
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| Boiling Point |
266.7ºC at 760 mmHg
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| Melting Point |
260ºC
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| Flash Point |
115.1ºC
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| LogP |
1.072
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
3
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| Rotatable Bond Count |
1
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| Heavy Atom Count |
11
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| Complexity |
174
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
PIVDNPNYIBGXPL-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C7H11NO2.ClH/c1-8-4-2-3-6(5-8)7(9)10;/h3H,2,4-5H2,1H3,(H,9,10);1H
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| Chemical Name |
1-methyl-3,6-dihydro-2H-pyridine-5-carboxylic acid;hydrochloride
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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 : ~100 mg/mL (~562.97 mM)
DMSO : ~25 mg/mL (~140.74 mM) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (14.07 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 25.0 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: ≥ 2.5 mg/mL (14.07 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 25.0 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: ≥ 2.5 mg/mL (14.07 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 | 5.6297 mL | 28.1484 mL | 56.2968 mL | |
| 5 mM | 1.1259 mL | 5.6297 mL | 11.2594 mL | |
| 10 mM | 0.5630 mL | 2.8148 mL | 5.6297 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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