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AP-III-a4 (ENOblock)

AP-III-a4 (ENOblock) is the first nonsubstrate analogue inhibitor of enolase with IC50 of 0.576 μM.

AP-III-a4 (ENOblock) Chemical Structure

AP-III-a4 (ENOblock) Chemical Structure

CAS No. 1177827-73-4

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AP-III-a4 (ENOblock) Related Products

Biological Activity

Description AP-III-a4 (ENOblock) is the first nonsubstrate analogue inhibitor of enolase with IC50 of 0.576 μM.
Targets
enolase [1]
(Cell-free assay)
0.576 μM
In vitro
In vitro In HCT116 cells, AP-III-a4 induces cell death under hypoxia, and inhibits cancer cell migration and invasion by down-regulation of AKT and Bcl-xL expression. In Huh7 hepatocytes and HEK kidney cells, AP-III-a4 induces glucose uptake and inhibits phosphoenolpyruvate carboxykinase (PEPCK) expression. [1]
Kinase Assay Enolase activity assay
A single unit of enolase is defined as the amount of enzyme that produces 1 μmol of phosphoenol pyruvate from phospho-D-glycerate/min in standard assay. Enolase activity assay is measured at 37°C by incubating pure enolase (3–9 U) in a buffer containing 50 mM imidazole-HCl (pH 6.8), 2.0 mM MgSO4 and 400 mM KCl in the absence or presence of ENOblock or NaF. The reaction is initiated by adding 1 μmol of 2-phospho-D-glycerate, and the OD is measured after 10 min of reaction time with a spectrophometer at 240 nm.
Cell Research Cell lines HCT116 cells
Concentrations 5 μM
Incubation Time 24 h
Method

3 x 105 HCT116 cells are seeded in a 6 well plate. 24 h later, cells are treated with compound of interest (with or without prior induction of hypoxia for 4 h) for 24 h using triplicate wells. Cells are then trypsinized and resuspended in 2.5 mL PBS. A 100 μL aliquot is taken for staining with 0.2% trypan blue solution and counted using a hemocytometer. 150 cells are counted and dead cells are classified as those that could not exclude trypan blue.
Experimental Result Images Methods Biomarkers Images PMID
Western blot p-AMPKα / AMPKα / p-ACC / ACC p-AKT / AKT / p-GSK3β / GSK3β / p-PRAS40 / PRAS40 30242159
In Vivo
In vivo In a HCT116-xenotransplanted zebrafish tumor xenograft model, AP-III-a4 (10 μM) inhibits cancer cell migration and invasion processes. In vivo, AP-III-a4 (10 μM) also causes downregulation of PEPCK expression and induction of glucose uptake, and inhibits adipogenesis and foam cell formation. [1]
Animal Research Animal Models HCT116-xenotransplanted zebrafish tumor xenograft model
Dosages 10 μM
Administration --

Chemical Information & Solubility

Molecular Weight 631.18 Formula

C31H44ClFN8O3
CAS No. 1177827-73-4 SDF Download AP-III-a4 (ENOblock) SDF
Smiles C1CCC(CC1)CNC2=NC(=NC(=N2)NCC3=CC=C(C=C3)F)NC4=CC=C(C=C4)CC(=O)NCCOCCOCCN.Cl
Storage (From the date of receipt)

In vitro
Batch:

DMSO : 100 mg/mL ( (158.43 mM) Moisture-absorbing DMSO reduces solubility. Please use fresh DMSO.)

Water : 100 mg/mL

Ethanol : 100 mg/mL


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Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

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