| Size | Price | Stock | Qty |
|---|---|---|---|
| 10mg |
|
||
| 25mg |
|
||
| 50mg |
|
||
| 100mg | |||
| 500mg | |||
| 1g | |||
| Other Sizes |
| Targets |
Acetylcholinesterase (AChE). AChE-IN-30 is a potent, small-molecule inhibitor of the enzyme AChE, which is responsible for the hydrolysis of the neurotransmitter acetylcholine. By inhibiting AChE, it increases the concentration and duration of action of acetylcholine in the synaptic cleft, thereby enhancing cholinergic neurotransmission. Additionally, it has a secondary cytoprotective effect by reducing ROS levels, which is independent of its AChE inhibition.
|
|---|---|
| ln Vitro |
AChE-IN-30 inhibits acetylcholinesterase activity in cell-free assays with an IC50 value of 4.4 microM. This demonstrates potent and direct inhibition of the target enzyme. It is an AChE inhibitor (antagonist). This level of potency is in the range that is considered drug-like for CNS (central nervous system) targets.
|
| ln Vivo |
AChE-IN-30 has neuroprotective activity and inhibits H2O2-induced apoptosis by suppressing intracellular ROS accumulation. While specific in vivo data is not provided, its mechanism (AChE inhibition + antioxidant) makes it a strong candidate for improving cognitive function and providing neuroprotection in animal models of Alzheimer's disease (AD). It would be evaluated in memory and learning tasks like the Morris water maze.
|
| Enzyme Assay |
AChE-IN-30 is a small-molecule acetylcholinesterase (AChE) inhibitor. For a cell-free AChE inhibition assay, a standard protocol uses the Ellman's method. Recombinant human AChE (or electric eel AChE) is diluted in assay buffer (0.1 M sodium phosphate, pH 8.0). Varying concentrations of AChE-IN-30 (0.1 nM-100 uM) are incubated with the enzyme for 15-30 minutes at 37degC. The reaction is then started by adding the substrates acetylthiocholine iodide (ATChI, 0.5 mM) and 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB, 0.33 mM). The production of the yellow 5-thio-2-nitrobenzoate anion is measured spectrophotometrically at 412 nm for 5-10 minutes. The IC50 is the concentration required to inhibit 50% of the enzyme's activity. For a reversible inhibition study, the enzyme is incubated with the compound and the inhibitor's mode of action (competitive/non-competitive) is determined using Lineweaver-Burk plots.
|
| Cell Assay |
For in vitro cellular studies, a neuronal cell line that expresses AChE, such as SH-SY5Y or PC12 cells, is used. Cells are seeded in 96-well plates (1×10⁴ cells/well). To induce an Alzheimer's-like state, cells are treated with amyloid-beta (Abeta1-42) oligomers (5-10 uM). AChE-IN-30 is then added at concentrations ranging from 1-100 uM. After 24-48 hours, AChE enzyme activity is measured in cell lysates using the Ellman's method. For neuroprotection, cells are treated with H2O2 (100-300 uM) to induce oxidative stress. AChE-IN-30 (1-50 uM) is co-incubated. Cell viability is measured by MTT assay. ROS levels are measured using the DCFH-DA fluorescent probe. Apoptosis is assessed by measuring caspase 3/7 activity or by flow cytometry using Annexin V/PI staining.
|
| Animal Protocol |
No specific in vivo animal study protocols are documented. For a standard Alzheimer's disease study, a typical protocol would involve using 8-12 month old APP/PS1 transgenic mice (amyloid model) or scopolamine-induced amnesia model in C57BL/6 mice. AChE-IN-30 would be administered daily via intraperitoneal (i.p.) injection or oral gavage at doses of 5-30 mg/kg for 2-4 weeks. The primary endpoint would be cognitive performance in the Morris water maze (learning and memory) or novel object recognition test. At the end of the study, brain tissues would be harvested for AChE activity measurement, analysis of oxidative stress markers (e.g., malondialdehyde, glutathione), and histopathology for amyloid plaque burden or neuronal loss. Blood-brain barrier penetration would be assessed by LC-MS analysis of compound levels in brain homogenates.
|
| ADME/Pharmacokinetics |
No pharmacokinetic data has been reported. As a small molecule with a molecular weight of 376.45 g/mol, and because it is designed as a CNS (central nervous system) drug, it is predicted to be able to cross the blood-brain barrier (BBB), which is a critical requirement for treating Alzheimer's disease. Its specific ADME (absorption, distribution, metabolism, excretion) profile has not been published. It is likely to be metabolized in the liver.
|
| Toxicity/Toxicokinetics |
No specific toxicity data has been reported. AChE inhibitors as a class are known to have a cholinergic toxicity profile at high doses. This can include side effects such as nausea, vomiting, diarrhea, muscle cramps, bradycardia (slow heart rate), and excessive salivation (SLUDGE syndrome). However, AChE-IN-30 is a research chemical and its in vivo toxicity profile has not been assessed in the literature available. Safety should be confirmed by the user.
|
| Additional Infomation |
AChE-IN-30 is a novel, small-molecule AChE inhibitor. It is being developed as a research tool to investigate both the cholinergic and oxidative stress components of Alzheimer's disease (AD). The dual mechanism of action (AChE inhibition + antioxidant) makes it a unique probe for studying multifactorial aspects of neurodegeneration. Its IUPAC name is not provided, but its molecular formula is C22H24N4O2. It is not approved for clinical use.
|
| Exact Mass |
376.19
|
|---|---|
| Elemental Analysis |
C, 70.19; H, 6.43; N, 14.88; O, 8.50
|
| CAS # |
2937454-22-1
|
| PubChem CID |
168459173
|
| Appearance |
Typically exists as solid at room temperature
|
| Hydrogen Bond Donor Count |
3
|
| Hydrogen Bond Acceptor Count |
6
|
| Rotatable Bond Count |
6
|
| Heavy Atom Count |
28
|
| Complexity |
610
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
C1CN(CCC1NC2=C(C(=O)C2=O)NC3=CC=C(C=C3)N)CC4=CC=CC=C4
|
| InChi Key |
RMNHWEFBTVCRRS-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C22H24N4O2/c23-16-6-8-17(9-7-16)24-19-20(22(28)21(19)27)25-18-10-12-26(13-11-18)14-15-4-2-1-3-5-15/h1-9,18,24-25H,10-14,23H2
|
| Chemical Name |
3-((4-aminophenyl)amino)-4-((1-benzylpiperidin-4-yl)amino)cyclobut-3-ene-1,2-dione
|
| Synonyms |
AChE-IN-30; AChE-IN30; AChE-IN 30
|
| HS Tariff Code |
2934.99.9001
|
| 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)
|
| 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
|
|---|---|
| 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.) |
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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
