Absorption, Distribution and Excretion
Oral administration - poor gastrointestinal absorption. Metabolism/Metabolites Plasma and Liver

Toxicity Summary
Abenidine is a cholinesterase, or acetylcholinesterase (AChE) inhibitor. Cholinesterase inhibitors (or "anticholinesterases") inhibit the activity of acetylcholinesterase. Because acetylcholinesterase plays a vital physiological role, chemicals that interfere with its activity are potent neurotoxins; even low doses can cause excessive salivation and lacrimation, followed by muscle spasms and ultimately death. Substances used in nerve gases and many pesticides have been shown to exert their effects by binding to serine residues at the active site of acetylcholinesterase, thus completely inhibiting the enzyme's activity. Acetylcholinesterase breaks down the neurotransmitter acetylcholine, which is released at the neuromuscular junction, causing muscle or organ relaxation. Inhibition of acetylcholinesterase results in the accumulation and sustained action of acetylcholine, leading to the continuous transmission of nerve impulses and an inability to stop muscle contractions. The most common acetylcholinesterase inhibitors are phosphorus-containing compounds designed to bind to the enzyme's active site. Its structural requirements include a phosphorus atom with two lipophilic groups, a leaving group (e.g., a halide or thiocyanate), and a terminal oxygen atom.

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Toxicity Data
LD₅₀ = 150 ± 44 mg/kg (oral in mice).

Ambenonium is a symmetrical oxalamide bisquaternary ammonium ion with an ethyl and a 2-chlorobenzyl group attached to its nitrogen atom. It is an EC 3.1.1.8 (cholinesterase) inhibitor. Ambenonium is a cholinesterase inhibitor used to treat myasthenia gravis. The mechanism of action of Ambenonium is as a cholinesterase inhibitor. Ambenonium is a bisquaternary ammonium alcohol with parasympathomimetic activity. Its positive charge allows it to act as an acetylcholinesterase inhibitor by binding to the anionic site of the active site of acetylcholinesterase, thereby preventing the breakdown of acetylcholine and producing indirect cholinergic effects on nicotine and muscarinic receptors. Ambenonium also has direct nicotine receptor agonist activity, enhancing cholinergic agonism at the neuromuscular junction. Ambenonium is only present in individuals who have used or taken this drug. It is a cholinesterase inhibitor used to treat myasthenia gravis. [Wikipedia] Ambecium chloride exerts its therapeutic effect on myasthenia gravis by competitively and reversibly inhibiting acetylcholinesterase. Myasthenia gravis is caused by the abnormal production of antibodies against acetylcholine receptors, which inhibits normal acetylcholine signaling (when acetylcholine binds to acetylcholine receptors on striated muscle fibers, it stimulates these muscle fibers to contract). Ambecium chloride reversibly binds to the anionic site of acetylcholinesterase, thereby blocking the binding site of acetylcholine, inhibiting acetylcholine hydrolysis, and enhancing cholinergic function through the accumulation of acetylcholine at cholinergic synapses. This, in turn, promotes the transmission of nerve impulses at the neuromuscular junction, thus effectively treating the disease.

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Drug Indications
Ambecium chloride is used to treat muscle weakness caused by muscle diseases (myasthenia gravis).

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Mechanism of Action
Ambecium chloride exerts its anti-myasthenia gravis effect by competitively and reversibly inhibiting acetylcholinesterase. Myasthenia gravis is a disease caused by the body's abnormal production of antibodies against acetylcholine receptors, which inhibits normal acetylcholine signaling (when acetylcholine binds to acetylcholine receptors on skeletal muscle fibers, it stimulates these fibers to contract). Ammonium chloride reversibly binds to the anionic site of acetylcholinesterase, thereby blocking the binding site of acetylcholine, inhibiting the hydrolysis of acetylcholine, and enhancing cholinergic function through the accumulation of acetylcholine at cholinergic synapses. Elevated acetylcholine levels can promote the transmission of nerve impulses at the neuromuscular junction.

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Pharmacodynamics
Ammonium chloride, similar to pyridostigmine and neostigmine, is used to treat muscle weakness and fatigue in patients with myasthenia gravis. It is speculated that ammonium chloride exerts its therapeutic effect by enhancing cholinergic function by inhibiting the hydrolysis of acetylcholine by acetylcholinesterase. Elevated acetylcholine levels produce peripheral effects because acetylcholine is also present in the brain and tends to cause excitatory effects. Glands receiving impulses from the parasympathetic nervous system are also stimulated in the same way. This is why elevated acetylcholine levels lead to a decrease in heart rate and an increase in salivation. Ampicillin is used less frequently than neostigmine or pyridostigmine, but it may be a better choice for patients with bromide allergies. Ampicillin produces fewer muscarinic side effects than neostigmine, but more than pyridostigmine.

C28H42N4O2CL2+2

537.56468

536.268

7648-98-8

2131

White to off-white solid powder

196-199 °C
196 - 199 °C

6.319

2

2

14

36

614

0

CC[N+](CCNC(C(NCC[N+](CC)(CC1=CC=CC=C1Cl)CC)=O)=O)(CC2=CC=CC=C2Cl)CC.[Cl-].[Cl-]

OMHBPUNFVFNHJK-UHFFFAOYSA-P

InChI=1S/C28H40Cl2N4O2/c1-5-33(6-2,21-23-13-9-11-15-25(23)29)19-17-31-27(35)28(36)32-18-20-34(7-3,8-4)22-24-14-10-12-16-26(24)30/h9-16H,5-8,17-22H2,1-4H3/p+2

(2-chlorophenyl)methyl-[2-[[2-[2-[(2-chlorophenyl)methyl-diethylazaniumyl]ethylamino]-2-oxoacetyl]amino]ethyl]-diethylazanium

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

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

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.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*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).

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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)]
*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.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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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).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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 (Please use freshly prepared in vivo formulations for optimal results.)