| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| 5mg |
|
||
| Other Sizes |
| Targets |
AEP-IN-2 targets asparagine endopeptidase (AEP, also known as legumain), a lysosomal cysteine protease that cleaves proteins after asparagine residues. In the context of Alzheimer's disease, AEP cleaves APP at specific sites to generate aggregation-prone Aβ fragments and cleaves tau to generate neurotoxic fragments that promote tau aggregation and neurofibrillary tangle formation. By inhibiting AEP, AEP-IN-2 prevents the generation of these pathogenic protein fragments, thereby reducing amyloid plaque formation and tau pathology.
|
|---|---|
| ln Vitro |
AEP-IN-2 shows potent in vitro activity as an AEP inhibitor, effectively blocking the cleavage of APP and tau by AEP in biochemical assays. The compound reduces the levels of Aβ40, Aβ42, and p-Tau in cellular models, demonstrating its ability to inhibit AEP-mediated protein processing. The compound is orally active, suggesting good permeability and stability in the gastrointestinal tract. The specific IC50 value for AEP inhibition has not been detailed in the available literature but is expected to be in the sub-micromolar range based on its reported activity.
|
| ln Vivo |
In vivo activity of AEP-IN-2 has been demonstrated in preclinical models of Alzheimer's disease. The compound reduces Aβ40, Aβ42, and p-Tau levels in animal models, consistent with its mechanism of AEP inhibition. Its oral bioavailability enables convenient administration via oral gavage, supporting its potential for chronic dosing in Alzheimer's disease patients. The reduction of both amyloid and tau pathologies suggests that AEP-IN-2 may have disease-modifying potential by targeting two key pathological hallmarks of Alzheimer's disease simultaneously.
|
| Enzyme Assay |
Non-cell-based enzyme assays for AEP-IN-2 typically involve measuring the inhibition of recombinant AEP activity using fluorogenic peptide substrates. A standard protocol includes incubating purified AEP enzyme with a fluorogenic substrate (such as Z-Ala-Ala-Asn-AMC) in assay buffer (pH 5.5–6.0) at 37°C. AEP-IN-2 is added at various concentrations (typically 0.001–100 μM), and the release of fluorescent AMC is monitored over time using a fluorescence plate reader. IC50 values are calculated from dose-response curves, and the mode of inhibition (competitive, non-competitive, or uncompetitive) can be determined by Lineweaver-Burk analysis.
|
| Cell Assay |
Cellular assays for AEP-IN-2 typically involve neuronal cell lines or primary neurons expressing APP and tau. A representative protocol includes culturing cells (e.g., SH-SY5Y or primary cortical neurons) in appropriate medium, treating with AEP-IN-2 at various concentrations (0.01–100 μM) for 24–72 hours, and measuring AEP activity using a cell-permeable fluorogenic substrate. Aβ40 and Aβ42 levels in the conditioned medium are quantified by ELISA, and p-Tau levels are assessed by Western blot using phosphorylation-specific antibodies. Cell viability is assessed to ensure that observed effects are not due to cytotoxicity.
|
| Animal Protocol |
In vivo animal studies with AEP-IN-2 are conducted in transgenic mouse models of Alzheimer's disease, such as APP/PS1 or 3xTg-AD mice. A typical protocol involves administering AEP-IN-2 via oral gavage at doses determined from pharmacokinetic studies (typically 10–50 mg/kg, once or twice daily) for several weeks or months. At the end of the treatment period, brain tissues are harvested for analysis of Aβ40, Aβ42, and p-Tau levels by ELISA or immunohistochemistry. Cognitive function is assessed using behavioral tests such as the Morris water maze or novel object recognition.
|
| ADME/Pharmacokinetics |
Pharmacokinetic properties of AEP-IN-2 have been characterized for oral administration. The compound has a molecular weight of 363.39 g/mol and is orally active, suggesting favorable bioavailability and permeability. The compound is stored as a powder at -20°C for up to 3 years and in solvent at -80°C for 1 year. Standard PK parameters including half-life, Cmax, and AUC can be determined from plasma and brain concentration-time profiles following oral administration. Brain penetration is a critical parameter for Alzheimer's disease therapeutics, and AEP-IN-2 is expected to cross the blood-brain barrier based on its oral activity in the CNS.
|
| Toxicity/Toxicokinetics |
As a research compound, AEP-IN-2 is not intended for human therapeutic use, and comprehensive toxicological data are limited to preclinical studies. Standard safety assessments would include cytotoxicity screening in neuronal and non-neuronal cell lines, hERG channel inhibition testing, and preliminary toxicology studies in animal models to determine maximum tolerated dose and identify potential target organs of toxicity. The compound is stored as a powder at -20°C for up to 3 years.
|
| References | |
| Additional Infomation |
AEP-IN-2 is a research tool for studying the role of asparagine endopeptidase in Alzheimer's disease pathogenesis. By targeting both amyloid and tau pathologies through AEP inhibition, this compound represents a promising approach for disease-modifying therapy. The compound is referenced in the primary literature for its ability to block AEP cleavage of APP and tau and reduce Aβ and p-Tau levels. AEP-IN-2 is suitable for both in vitro and in vivo studies, providing researchers with a tool to investigate the therapeutic potential of AEP inhibition in Alzheimer's disease.
|
| Exact Mass |
363.111
|
|---|---|
| CAS # |
2565572-83-8
|
| PubChem CID |
155341905
|
| Appearance |
Light yellow to yellow solid powder
|
| Hydrogen Bond Donor Count |
3
|
| Hydrogen Bond Acceptor Count |
11
|
| Rotatable Bond Count |
5
|
| Heavy Atom Count |
25
|
| Complexity |
445
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
CNC1=NN=C(S1)NC2=CC(=C(C3=NON=C23)N4CCOCC4)CO
|
| InChi Key |
BSMQYCJYUKHJAX-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C14H17N7O3S/c1-15-13-17-18-14(25-13)16-9-6-8(7-22)12(11-10(9)19-24-20-11)21-2-4-23-5-3-21/h6,22H,2-5,7H2,1H3,(H,15,17)(H,16,18)
|
| Chemical Name |
[7-[[5-(methylamino)-1,3,4-thiadiazol-2-yl]amino]-4-morpholin-4-yl-2,1,3-benzoxadiazol-5-yl]methanol
|
| 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) |
Typically soluble in DMSO (e.g. 10 mM)
|
|---|---|
| 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
