| Targets |
AChE-IN-65 targets acetylcholinesterase (AChE), the enzyme responsible for hydrolyzing the neurotransmitter acetylcholine in the synaptic cleft. As a mixed inhibitor, it binds to both the free enzyme and the enzyme-substrate complex, reducing the rate of acetylcholine breakdown and thereby increasing acetylcholine availability at cholinergic synapses. This mechanism is relevant to Alzheimer's disease and other neurodegenerative disorders characterized by cholinergic deficit.
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| ln Vitro |
In vitro, AChE-IN-65 acts as a mixed inhibitor of Electrophorus electricus AChE, with a Ki value of 556.4 microM. This indicates relatively weak inhibitory potency compared to other AChE inhibitors. No cellular activity data (e.g., IC50 on cell lines) or detailed enzyme kinetic parameters (Vmax, Km) are reported in the search results beyond the Ki value. The compound is a tool for studying mixed inhibition mechanisms in cholinesterase research.
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| ln Vivo |
No in vivo activity data for AChE-IN-65 are reported in the search results. As a weak AChE inhibitor (Ki = 556.4 microM), it would likely require high doses to achieve significant AChE inhibition in animal models. It could potentially be evaluated in rodent models of Alzheimer's disease (e.g., scopolamine-induced amnesia models) or other cholinergic disorders, but no such data are currently available.
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| Enzyme Assay |
The binding affinity of AChE-IN-65 to acetylcholinesterase is measured using Ellman‘s colorimetric assay. Electrophorus electricus AChE is incubated with the compound at graded concentrations (1 microM-10 mM) in phosphate buffer (pH 8.0) for 10 min. Acetylthiocholine iodide (substrate) and DTNB (Ellman's reagent) are then added, and absorbance at 412 nm is measured spectrophotometrically. The inhibition constant (Ki) of 556.4 microM is calculated from Lineweaver-Burk plots or Dixon plots by varying both inhibitor and substrate concentrations. The mixed inhibition pattern is confirmed by observing changes in both Km and Vmax.
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| Cell Assay |
No cellular experiments for AChE-IN-65 are described in the search results. In a typical AChE inhibitor cellular assay, neuronal cell lines (e.g., SH-SY5Y, IMR-32, PC12) would be seeded in 96-well plates and treated with AChE-IN-65 at graded concentrations (1-1000 microM) for 24-72 h. Intracellular AChE activity would be measured using a commercial fluorescence-based kit, and cell viability would be assessed by MTT assay to evaluate cytotoxicity. However, no such data are reported for this compound.
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| Animal Protocol |
No animal experiments for AChE-IN-65 are described in the search results. For in vivo evaluation, male Sprague-Dawley rats or C57BL/6 mice would be administered AChE-IN-65 intraperitoneally or orally at doses of 10-200 mg/kg. After 30-120 min, animals would be sacrificed, and brain (cortex, hippocampus) and blood samples would be collected. AChE activity in brain homogenates and erythrocytes would be measured using Ellman's method. Cognitive effects could be assessed using the Morris water maze or passive avoidance test in scopolamine-induced amnesia models.
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| ADME/Pharmacokinetics |
AChE-IN-65 (C1₅H₈N2O₆, MW = 312.23, CAS: 98704-22-4) is a solid powder. For storage, the compound should be kept at -20degC in sealed containers, away from moisture and light. For in vitro use, stock solutions in DMSO should be stored at -80degC for up to 6 months or at -20degC for up to 1 month (sealed, away from moisture and light). Solubility and in vivo formulation details are not reported in the search results.
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| Toxicity/Toxicokinetics |
No specific toxicity data for AChE-IN-65 are reported in the search results. As a research-grade compound, it is not intended for human use. Standard laboratory safety precautions, including the use of personal protective equipment (gloves, lab coat, goggles), should be followed during handling. No LD50 or formal toxicology study results are provided in the available sources.
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| References | |
| Additional Infomation |
AChE-IN-65 (Compound DABA_1) is a research-grade mixed inhibitor of acetylcholinesterase with weak potency (Ki = 556.4 microM). It is not a drug and has not entered clinical trials or received regulatory approval for any indication. The compound serves as a chemical tool for studying cholinesterase inhibition mechanisms, particularly mixed-type inhibition, and for exploring the structure-activity relationships of AChE inhibitors. It may be used in biochemical assays as a reference compound for screening more potent AChE inhibitors.
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| CAS # |
98704-22-4
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| Appearance |
Solid powder
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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 (e.g. under nitrogen), avoid exposure to moisture and light. |
| 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 : 100 mg/mL (320.28 mM; with sonication)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (8.01 mM)(Saturation unknown) in 10% DMSO 40% PEG300 5% Tween-80 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, add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix well; then add 50 μL Tween-80 to the above system and mix well; then add 450 μL saline to make it 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 (8.01 mM)(saturation unknown) in 10% DMSO 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution, add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix. Take μL of DMSO stock solution, add μL . μL , mix until clear, add μL Tween 80, mix until clear, and add μL of saline. Dissolve 0.9 g of sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear and transparent saline solution. If the continuous administration period exceeds half a month, please choose this solution with caution. Please ensure that the first step stock solution is dissolved to a clear state, and add the co-solvent from left to right. You can use ultrasonic heating (ultrasonic cleaning instrument, recommended frequency 20-40 kHz), vortex blowing and other methods to assist dissolution. (Please use freshly prepared in vivo formulations for optimal results.) |
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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