| Size | Price | Stock | Qty |
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| 1mg |
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| Other Sizes |
| Targets |
AChRs:acetylcholine receptors
Asoxime targets organophosphate‑inhibited acetylcholinesterase (AChE) at the cholinergic synapse. The oxime group of asoxime binds to the phosphyl group attached to the active site serine residue of AChE and performs a nucleophilic attack, cleaving the organophosphate‑enzyme bond and reactivating the enzyme. Asoxime has a broader spectrum of reactivation against different organophosphates compared to pralidoxime (2‑PAM) and is particularly effective against soman‑inhibited AChE. |
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| ln Vitro |
Specific in vitro activity data for Asoxime-d4 are not available, as it is utilized as a tracer or analytical standard. However, the parent compound, Asoxime dichloride (HI-6), acts as an antagonist at acetylcholine receptors, including the α7 nicotinic acetylcholine receptor (α7 nAChR). This activity underlies its pharmacological effects and its use in research models .
Drug compounds have included stable heavy isotopes of carbon, hydrogen, and other elements, mostly as tracers for quantification throughout the drug development process. Due to its potential to alter the pharmacokinetic and metabolic characteristics of medications, deuteration has drawn attention[1]. In cell‑free enzymatic assays, asoxime (non‑deuterated) reactivates organophosphate‑inhibited human recombinant AChE with a rate constant (kr) typically in the range of 10⁴‑10⁵ M-¹·min-¹, depending on the specific organophosphate. The reactivation potency (EC50) is in the low micromolar to sub‑micromolar range. Asoxime has a higher affinity for the AChE active site than pralidoxime and can penetrate the blood‑brain barrier to a limited extent. The deuterated analogue has identical reactivation kinetics. |
| ln Vivo |
Specific in vivo activity data for Asoxime-d4 are not available. The in vivo profile of the parent compound, Asoxime (HI-6), is well-established as an effective antidote for organophosphate poisoning, such as from nerve agents. Its role as a cholinergic antagonist forms the basis for its therapeutic application in modulating the nervous system .
In vivo, asoxime (HI‑6) is the most effective broad‑spectrum reactivator of AChE inhibited by various nerve agents. In rodent models of organophosphate poisoning (e.g., sarin, VX, soman), intravenous or intramuscular administration of asoxime (10‑50 mg/kg) in combination with atropine (an anti‑muscarinic agent) and an anticonvulsant (e.g., diazepam) significantly increases survival and reduces neurological deficits. Asoxime reactivates AChE in peripheral tissues (e.g., blood, diaphragm) and the brain, restoring normal cholinergic neurotransmission. The deuterated analogue is not used therapeutically but is used to quantify asoxime levels in PK studies. |
| Enzyme Assay |
A standard in vitro AChE reactivation assay: Human recombinant AChE (or electric eel AChE) is incubated with an organophosphate (e.g., sarin, VX, paraoxon, 1‑10 uM) for 30‑60 min at 25degC to achieve >95% inhibition of enzyme activity. Excess organophosphate is removed by dilution or dialysis. The inhibited enzyme is then incubated with asoxime‑d4 dichloride (0.1‑1000 uM) in phosphate buffer (50 mM, pH 7.4) at 25degC or 37degC for various time points (0‑60 min). Residual AChE activity is measured using a colorimetric assay (Ellman's method), in which the hydrolysis of acetylthiocholine (ATCh, 1 mM) is monitored at 412 nm. The reactivation rate constant (kr) and the dissociation constant (Kd) are calculated using established kinetic models. The total AChE activity is determined by parallel incubations with a known reactivator.
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| Cell Assay |
A general cellular AChE reactivation assay: Human neuroblastoma cells (e.g., SH‑SY5Y) or primary cortical neurons are treated with an organophosphate (e.g., paraoxon, 10‑100 uM) for 30 min to inhibit cellular AChE. Cells are washed to remove excess organophosphate, then treated with asoxime‑d4 dichloride (1‑100 uM) for 30‑60 min. Cells are lysed, and AChE activity is measured in the lysate using the Ellman's method. Protein concentration is determined by BCA assay, and AChE activity is normalised to total protein. Percent reactivation is calculated relative to non‑inhibited control cells. Cytotoxicity of the organophosphate and asoxime is assessed by LDH release or MTT assay.
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| Animal Protocol |
A general animal model of organophosphate poisoning: Male Sprague‑Dawley rats (250‑300 g, n=6/group) are injected subcutaneously or intraperitoneally with a lethal or sub‑lethal dose of an organophosphate (e.g., sarin, VX, soman, paraoxon) at 0.8‑1.5× LD50. One minute after organophosphate exposure, animals receive atropine sulphate (10‑20 mg/kg, i.p.) and asoxime (10‑50 mg/kg, i.m. or i.v.). A third agent, such as diazepam (10 mg/kg, i.p.) or an anticonvulsant, may be administered to control seizures. Survival is monitored for 24‑72 h. For PK studies, asoxime‑d4 is administered as a tracer or internal standard. At various time points (0‑8 h), blood samples are collected, and plasma is separated. Asoxime concentrations are determined by LC‑MS/MS using asoxime‑d4 as the internal standard. Brain tissue is also collected for AChE activity measurement (Ellman's assay) and quantification of asoxime levels. The effective dose (ED50) for reactivation and the protection ratio (PR) are calculated.
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| ADME/Pharmacokinetics |
As a deuterium-labeled compound, the pharmacokinetic properties of Asoxime-d4 are likely to be similar to those of the parent drug, Asoxime (HI-6), but with key differences due to the kinetic isotope effect. Deuteration can alter the rate of metabolic breakdown, potentially leading to increased metabolic stability, a longer half-life, and a reduced rate of clearance compared to the non-deuterated compound. These properties make it particularly suitable for use as an internal standard in bioanalytical methods .
As a deuterated analogue of asoxime, asoxime‑d4 dichloride has identical physicochemical and pharmacokinetic properties to the non‑deuterated compound. Asoxime has a short plasma half‑life (t1/2 30‑60 min in rodents) due to rapid renal excretion and metabolism. It has a low volume of distribution (~0.3‑0.5 L/kg) and is not highly plasma protein bound (<30%). Asoxime is a quaternary ammonium compound, which limits its penetration of the blood‑brain barrier (BBB); however, sufficient amounts enter the brain to reactivate AChE in the central nervous system. The compound is excreted unchanged in the urine (60‑80% within 4 h). The deuterated form is used as an internal standard to improve the accuracy of LC‑MS/MS quantitation. |
| Toxicity/Toxicokinetics |
No specific toxicity data for Asoxime-d4 dichloride (HI-6-d4) were found in the available resources. Its use is strictly for research purposes. For the non-deuterated parent compound, Asoxime (HI-6), its safety profile is derived from its use as a pharmaceutical agent. The deuterated version is intended for laboratory use and is not a finished pharmaceutical product for human or veterinary use .
Asoxime is generally well‑tolerated in animals at therapeutic doses (10‑50 mg/kg, i.m./i.v.). At very high doses (>100 mg/kg), mild neuromuscular blockade, hypotension and injection site pain may occur. In humans, HI‑6 has been evaluated in clinical trials and is considered safe, with mild adverse events including transient increases in liver enzymes, headache, dizziness and injection site pain. As a research‑grade isotopologue, asoxime‑d4 is for research use only and is not intended for human therapeutic administration. Standard safety precautions (gloves, lab coat) should be used when handling. |
| References | |
| Additional Infomation |
Asoxime-d4 dichloride (HI-6-d4) is the deuterium-labeled form of Asoxime dichloride (HI-6). It is a stable isotope-labeled compound intended primarily for research use, such as acting as an internal standard in liquid chromatography-mass spectrometry (LC-MS) for the quantification of HI-6 in biological samples. The parent compound, Asoxime (HI-6), is an antagonist of acetylcholine receptors (AChRs), specifically targeting the nicotinic receptor, α7 nAChR. Its research applications include modulating immune responses and improving vaccination efficacy in the nervous system
Asoxime (HI‑6) is one of the most promising broad‑spectrum oxime reactivators for nerve agent poisoning. It is part of the standard military nerve agent antidote kits in several countries (e.g., Canada, Sweden, Switzerland). The deuterated form is used as an internal standard for the quantitative bioanalysis of asoxime in pharmacokinetic, forensic and toxicological studies. Asoxime‑d4 is supplied as a dichloride salt (the same salt form as the parent drug) to ensure identical solubility and handling properties. For research use only; not for human therapeutic use. |
| Molecular Formula |
C14H12D4CL2N4O3
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|---|---|
| Molecular Weight |
363.23
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| Exact Mass |
362.08505
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| Related CAS # |
Asoxime dichloride;34433-31-3
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| PubChem CID |
136257807
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| Appearance |
White to off-white solid powder
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
6
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| Heavy Atom Count |
23
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| Complexity |
356
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| Defined Atom Stereocenter Count |
0
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| SMILES |
[2H]C1=C([N+](=C(C(=C1C(=O)N)[2H])[2H])COC[N+]2=CC=CC=C2/C=N/O)[2H].[Cl-].[Cl-]
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| InChi Key |
QELSIJXWEROXOE-FFVJOBTLSA-N
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| InChi Code |
InChI=1S/C14H14N4O3.2ClH/c15-14(19)12-4-7-17(8-5-12)10-21-11-18-6-2-1-3-13(18)9-16-20;;/h1-9H,10-11H2,(H-,15,19);2*1H/i4D,5D,7D,8D;;
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| Chemical Name |
2,3,5,6-tetradeuterio-1-[[2-[(E)-hydroxyiminomethyl]pyridin-1-ium-1-yl]methoxymethyl]pyridin-1-ium-4-carboxamide dichloride
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| Synonyms |
Asoxime-d4 (dichloride); Asoxime-d4 Chloride;
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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 Note: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| 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 | 2.7531 mL | 13.7654 mL | 27.5308 mL | |
| 5 mM | 0.5506 mL | 2.7531 mL | 5.5062 mL | |
| 10 mM | 0.2753 mL | 1.3765 mL | 2.7531 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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