AJ-4
AJ-4 is an orally active, blood-brain barrier-permeable GSK-3β/AChE inhibitor, with an IC50 of 4.7 nM against GSK-3β and 2.97 μM against AChE. AJ-4 increases the level of phosphorylated GSK-3β and reduces the expression of Amyloid-β and phosphorylated Tau. AJ-4 improves scopolamine-induced learning and memory impairment and alleviates hippocampal neuron damage in AD mice. AJ-4 can be used for the research of Alzheimer's disease.
For research use only. We do not sell to patients.
- Formula: C23H21Cl3N4O5S
- Molecular Weight:571.86
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Storage:
Please store the product under the recommended conditions in the Certificate of Analysis.
Biological Activity
Description
IC50 & Target
[1]|
GSK-3β 4.7 nM (IC50) |
AChE 2.97 μM (IC50) |
In Vitro
AJ-4 (10 μM; 24 h) significantly restores the viability of SH-SY5Y cells damaged by Aβ25-35[1].
AJ-4 (5-20 μM; 24 h) regulates key Alzheimer's disease (AD)-related proteins in Aβ25-35-damaged SH-SY5Y cells in a concentration-dependent manner, with the specific mechanism involving upregulation of p-GSK-3β and downregulation of APP and p-Tau[1].
AJ-4 (5-20 μM; 24 h) concentration-dependently inhibits Aβ25-35-induced secretion of TNF-α and IL-1β in SH-SY5Y cells, exhibiting anti-inflammatory activity[1].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only. Further protocols information, click here.
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Cell Line:Aβ25-35-injured SH-SY5Y human neuroblastoma cells
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Concentration:10 μM
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Incubation Time:24 h
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Result:Restored cell viability from 54.3% (Aβ25-35-injured level) to 98.98%.
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Cell Line:Aβ25-35-stimulated SH-SY5Y human neuroblastoma cells
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Concentration:5, 10, 20 μM
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Incubation Time:24 h
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Result:Concentration-dependently reduced Aβ25-35-upregulated TNF-α and IL-1β levels.
Showed the highest reduction of pro-inflammatory cytokines at 20 μM.
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Cell Line:Aβ25-35-injured SH-SY5Y human neuroblastoma cells
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Concentration:5, 10, 20 μM
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Incubation Time:24 h
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Result:Concentration-dependently downregulated APP and phosphorylated Tau (p-Tau) levels relative to the Aβ25-35-injured model group.
Concentration-dependently upregulated phosphorylated GSK-3β (p-GSK-3β) levels relative to the Aβ25-35-injured model group.
Parmacokinetics
In Vivo
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
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Animal Model:Kunming mice (male, scopolamine-induced AD-like behavior)[1]
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Dosage:0.5 mg/kg; 1.0 mg/kg; 2.0 mg/kg
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Administration:p.o.; daily; 18 days
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Result:Reduced orientation navigation escape latency in a dose-dependent manner starting from day 3 of training.
Increased the number of virtual platform crossings relative to the model group at 0.5 mg/kg.
Increased both the percentage of movement distance in the target quadrant and the number of virtual platform crossings relative to the model group at 1.0 mg/kg and 2.0 mg/kg.
Significantly downregulated APP and phosphorylated Tau (p-Tau) expression, while significantly upregulated phosphorylated GSK-3β (p-GSK-3β) expression relative to the model group at all tested doses.
Preserved clear hippocampal histomorphology, with only slight neuronal pyknosis and hyperchromasia limited to CA1 and DG regions at 1.0 mg/kg and 2.0 mg/kg.
Resulted in intact hippocampal structure with only sporadic pyknotic neurons in the CA3 region at 2.0 mg/kg.
Chemical Information
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Molecular Weight 571.86
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Formula C23H21Cl3N4O5S
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SMILES
C=CC(NC1=C(C=CC(N2SC(N(C2=O)CC3=CC=C(C=C3)OC(N(CCCl)CCCl)=O)=O)=C1)Cl)=O
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Shipping
Room temperature in continental US; may vary elsewhere.
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Storage
Please store the product under the recommended conditions in the Certificate of Analysis.
Protocols
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RNA extraction experimental
By lysing cells, releasing RNA, and removing impurities such as proteins and DNA, high-purity RNA products are finally obtained. The commonly used traditional method is the guanidine isothiocyanate/phenol/chloroform method (Trizol), which is suitable for a variety of animal materials including animal tissues, microorganisms, cultured cells, etc., and most plant materials.
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Protocol for Shuttle Box Test (TDPA)
The Shuttle Box Test for TDPA, or temporally dissociated passive avoidance, measures hippocampus-dependent associative learning by testing whether a rodent avoids entering a dark compartment that was previously paired with foot shock after a temporal delay between dark-compartment entry and shock delivery. The main behavioral readout is crossover or step-through latency from the light chamber into the dark chamber; increased latency across training or retention trials reflects learned avoidance memory rather than motor performance alone.
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Cell Cytotoxicity Assay
Cytotoxicity assays are usually based on the assessment of cell membrane damage, which can also be indirectly detected by measuring cell viability. Detection methods include MTT assay, CKK-8 assay, LDH assay and ATP assay, etc.
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Protocol for Water Maze
The Morris Water Maze is a rodent spatial learning and memory assay in which a mouse or rat swims in opaque water to find an escape platform; in the hidden-platform version, the animal cannot see the platform and must use distal extra-maze cues to learn its fixed spatial location. The assay primarily measures hippocampus-dependent spatial learning during acquisition trials and spatial reference memory during probe trials after platform removal; readouts include escape latency, swim path length, swim speed, quadrant occupancy, platform-site crossings, and proximity to the former platform location.
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Amyloid: Congo Red Amyloid Staining
Congo red amyloid staining is a histochemical method used to detect extracellular amyloid deposits in tissue sections based on the affinity of Congo red dye for β-pleated sheet-rich protein aggregates. When bound to amyloid, Congo red produces characteristic apple-green birefringence under polarized light microscopy, which is widely regarded as a diagnostic feature of amyloid deposition in histopathology. The diagnostic principle relies on the combination of dye binding (congophilia) and optical anisotropy under polarized illumination, which distinguishes amyloid from most non-amyloid eosinophilic extracellular deposits in routine histological evaluation. Amyloid identification by Congo red staining remains a cornerstone in diagnostic pathology despite the availability of adjunct methods such as immunohistochemistry and mass spectrometry, particularly because of its ability to localize deposits directly within tissue architecture. The specificity of Congo red-positive deposits is incre
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Alzheimer’s Disease Modeling
Alzheimer’s Disease (AD) is a neurodegenerative disorder characterized by a progressive decline in cognitive functions and loss of specific types of neurons and synapses. Alzheimer's symptoms can be simulated in mice by injecting drugs (such as Aβ) or genetically modified.
Purity & Documentation
References
Calculators
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)