| Size | Price | Stock | Qty |
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| Targets |
D4.4 receptor ( Ki = 1.5 nM ); D4.2 receptor ( EC50 = 1.9 nM ); D4.7 receptor ( EC50 = 1.6 nM )
A-381393 targets dopamine D4 receptor. Binding affinity (Ki) for human D4.4 receptor: 1.5 ± 0.1 nM (using [³H]-spiperone competition binding assay). Selectivity >4000-fold over D1, D2L, D3, D5 dopamine receptors. Affinity for D4.2: Ki = 1.9 nM; for D4.7: Ki = 1.6 nM. Also shows moderate affinity for 5-HT2A receptor: Ki = 370 nM. No significant affinity (Ki > 1 μM) for >70 other receptors and channels, except 5-HT2A. [1] |
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| ln Vitro |
A-381393, with Kis of 1365, 8600, 2044, 1912, and 2962 nM, respectively, shows weak affinity at 5-HT1A, Sigma 2, Adenoceptor α1A, Adenoceptor α2C, and Histamine H1[1].
A-381393 potently inhibits dopamine-induced calcium flux in human D4.4 receptor-expressing cells. In FLIPR calcium flux assay, it inhibits the response to 1 μM dopamine with an IC50 of 8 nM. It shows no significant intrinsic agonist activity up to 10 μM. In contrast, L-745870 exhibits partial agonist activity (up to 20% of dopamine response at 10 μM). A-381393 does not affect dopamine responses in human D2L or D3 receptor calcium flux assays, demonstrating functional selectivity. In GTP-γ-S binding assay using human D4.2 membranes, A-381393 (100 and 1000 nM) has no intrinsic activity alone, but inhibits the response to suboptimal concentration (100 nM) of dopamine or the selective D4 agonist PD168077. Schild analysis indicates competitive antagonism with pA2 = 7.96 (corresponding to 11 nM). [1] |
| ln Vivo |
A-381393 inhibits penile erection induced by the selective D4 agonist PD168077 in conscious rats. PD168077 (0.3 μmol/kg, s.c.) induces 75% incidence of penile erection. Pretreatment with A-381393 (30 μmol/kg, s.c.) significantly reduces the incidence to 48% (p < 0.05). Observations are made for 60 min after dosing. [1]
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| Enzyme Assay |
Radioligand binding assays: Membranes from HEK-293 cells expressing recombinant human D4.4 receptor are incubated with 0.1 nM [³H]-spiperone in buffer containing 50 mM Tris-HCl (pH 7.4), 5 mM KCl, 120 mM NaCl, 5 mM MgCl₂, and 1 mM EDTA at room temperature for 2 h. Non‑specific binding is determined using 10 μM haloperidol. The reaction is terminated by rapid filtration through GF/B filters, and radioactivity is measured by scintillation counting. IC50 values are converted to Ki using the Cheng‑Prusoff equation. Similar procedures are used for D4.2, D4.7, D2L, D3, and other receptor binding assays. [1]
GTP‑Eu binding assay (DELFIA GTP‑binding kit): Membranes containing human D4.2 receptor (8 μg) are incubated with varying concentrations of ligand (0.1–10,000 nM) in buffer containing 50 mM HEPES (pH 7.4), 10 mM MgCl₂, 25 mM NaCl, 100 μg/ml saponin, 1 μM GDP, and 10 nM GTP‑Eu at room temperature for 40 min. Non‑specific binding is measured in the presence of 100 μM GTP‑γS. The reaction is terminated by rapid filtration, and filters are washed three times with ice‑cold wash buffer. Fluorescence is measured using a time‑resolved fluorometer. Percent stimulation is calculated relative to 10 μM dopamine‑stimulated binding. [1] |
| Cell Assay |
FLIPR calcium flux assay: Cells expressing human D4.4 receptor together with a chimeric G protein (Gzq₀₅) are plated into 96‑well black‑wall/clear‑bottom plates at 20,000 cells per well. After 2 days, culture medium is replaced with 0.1 ml DPBS containing 0.04% Pluronic F‑127 and 4 μM Fluo‑4, a fluorescent calcium indicator dye, and incubated for 1 h at room temperature. Cells are washed four times with DPBS, and 150 μl DPBS is added. Using a fluorometric imaging plate reader (FLIPR‑384), 50 μl of compound solution is transferred to the cells, and fluorescence is read for 3 min (every second for the first minute, then every 5 s for the next 2 min). Data are normalized to the response of 1 μM dopamine (or 10 μM dopamine in some experiments) for antagonist assays, or to 10 μM dopamine for intrinsic activity assessment. [1]
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| Animal Protocol |
Rat penile erection assay: Male adult Wistar rats (approx. 300 g) are used. On test day, animals acclimate to a dimly lit room with red light for 1 h before experiment. A-381393 is freshly prepared and administered subcutaneously (s.c.) into the back neck area above the left scapula (injection volume 1 ml/kg). For blockade experiments, rats receive A-381393 (30 μmol/kg, s.c.) before PD168077 (0.3 μmol/kg, s.c.). Immediately after injection, rats are placed individually into transparent Plexiglas cages (20×30×30 cm) with mirrors placed behind and under for observation. Penile erection episodes are recorded by direct observation for 60 min. An erection is defined as repeated pelvic thrusts followed by an upright position, emergence of an engorged penis, and grooming. Incidence (%) is the percentage of animals showing one or more erections. [1]
Pharmacokinetic study: Male Sprague‑Dawley rats (approx. 250 g, n=3/group) receive A-381393 at 1 mg/kg via intravenous (i.v.), subcutaneous (s.c.), or intraperitoneal (i.p.) routes. The compound is dissolved in isotonic dextrose (D5W) containing 10% ethanol, at appropriate concentrations for a 1 ml/kg dose volume. Serial blood samples are collected from a tail vein. At selected time points, rats are euthanized with CO₂, exsanguinated by cardiac puncture, and brains are removed. For brain penetration study, rats receive s.c. doses of 0.1, 1, or 10 mmol/kg, and are sacrificed at various time points (5‑60 min). Brain tissue is homogenized with two volumes of water. A-381393 is extracted from plasma and brain homogenate by liquid‑liquid extraction with ethyl acetate:hexane (1:1) at alkaline pH. The organic phase is evaporated, and the residue is reconstituted in mobile phase for HPLC‑MS/MS analysis (reverse phase C18 column, 0.1% TFA:acetonitrile 55:45, flow rate 0.4 ml/min). [1] |
| ADME/Pharmacokinetics |
Brain penetration: After s.c. administration of A-381393 at doses of 0.1, 1, or 10 mmol/kg to rats, plasma and brain levels increase proportionally with dose. At 10 mmol/kg s.c., plasma level reaches ~1200 ng/ml and brain level reaches ~500 ng/g.
Following s.c. administration (1 mg/kg) in rats, plasma Cmax = 34.3 ± 4.7 ng/ml. Following i.p. administration (1 mg/kg), plasma Cmax = 50.7 ± 17.1 ng/ml. Elimination half‑life: After i.v. administration (1 mg/kg), plasma t₁/₂ is 18 ± 6 min (calculated from 5 min to 1 h time points), indicating rapid metabolism in rats. A-381393 is not orally bioavailable. [1] |
| References | |
| Additional Infomation |
A-381393 (compound 33 in Cowart et al., 2004) is a novel, potent, and selective dopamine D4 receptor antagonist that lacks significant intrinsic agonist activity. It is identified using a functional calcium flux assay to distinguish agonist from antagonist activities. Compared to clozapine and L‑745870, A-381393 exhibits better receptor selectivity, with no affinity up to 10 μM for a panel of more than 70 receptors and channels, except moderate affinity for 5‑HT2A (Ki = 370 nM). Unlike L‑745870, A-381393 does not show partial agonist activity in D4 receptor assays. It is a valuable research tool for studying dopamine D4 receptors both in vitro and in vivo, particularly for investigating the role of D4 receptors in penile erection, cognition, attention deficit disorder, and schizophrenia. [1]
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| Molecular Formula |
C20H24N4
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|---|---|
| Molecular Weight |
320.431
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| Exact Mass |
320.2
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| Elemental Analysis |
C, 74.97; H, 7.55; N, 17.48
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| CAS # |
726174-00-1
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| PubChem CID |
11301655
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| Appearance |
Solid powder
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| LogP |
3.7
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
3
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| Rotatable Bond Count |
3
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| Heavy Atom Count |
24
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| Complexity |
406
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CC1=C(C=C(C=C1)N2CCN(CC2)CC3=NC4=CC=CC=C4N3)C
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| InChi Key |
SAQMCVDGOIRQTC-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C20H24N4/c1-15-7-8-17(13-16(15)2)24-11-9-23(10-12-24)14-20-21-18-5-3-4-6-19(18)22-20/h3-8,13H,9-12,14H2,1-2H3,(H,21,22)
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| Chemical Name |
2-[[4-(3,4-dimethylphenyl)piperazin-1-yl]methyl]-1H-benzimidazole
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| Synonyms |
A 381393; A381393; A-381393
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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: ~51.7 mg/mL (~161.3 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.58 mg/mL (8.05 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 25.8 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: ≥ 2.58 mg/mL (8.05 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 25.8 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: ≥ 2.58 mg/mL (8.05 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 | 3.1208 mL | 15.6040 mL | 31.2081 mL | |
| 5 mM | 0.6242 mL | 3.1208 mL | 6.2416 mL | |
| 10 mM | 0.3121 mL | 1.5604 mL | 3.1208 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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