AP-III-a4 (ENOblock)

Catalog No.S7443 Batch:S744301

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Technical Data

Formula

C31H44ClFN8O3
Molecular Weight 631.18 CAS No. 1177827-73-4
Solubility (25°C)* In vitro DMSO 100 mg/mL (158.43 mM)
Water 100 mg/mL (158.43 mM)
Ethanol 100 mg/mL (158.43 mM)
* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.
* Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

Preparing Stock Solutions

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 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]
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]

Protocol (from reference)

Kinase Assay:

[1]
  • 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 Assay:

[1]
  • 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.
Animal Study:

[1]
  • Animal Models

    HCT116-xenotransplanted zebrafish tumor xenograft model

  • Dosages

    10 μM

  • Administration

    --

Selleck's AP-III-a4 (ENOblock) has been cited by 9 publications

Mediation of PKM2-dependent glycolytic and non-glycolytic pathways by ENO2 in head and neck cancer development [ J Exp Clin Cancer Res, 2023, 42(1):1] PubMed: 36588153
A temperature-regulated circuit for feeding behavior [ Nat Commun, 2022, 13(1):4229] PubMed: 35869064
A temperature-regulated circuit for feeding behavior [ Nat Commun, 2022, 13(1):4229] PubMed: 35869064
The essential roles of m6A RNA modification to stimulate ENO1-dependent glycolysis and tumorigenesis in lung adenocarcinoma [ J Exp Clin Cancer Res, 2022, 41(1):36] PubMed: 35078505
Amoxicillin Haptenation of α-Enolase is Modulated by Active Site Occupancy and Acetylation [ Front Pharmacol, 2021, 12:807742] PubMed: 35095517
Preclinical validation of Alpha-Enolase (ENO1) as a novel immunometabolic target in multiple myeloma. [ Oncogene, 2020, 39(13):2786-2796] PubMed: 32024967
FGFRL1 affects chemoresistance of small-cell lung cancer by modulating the PI3K/Akt pathway via ENO1. [ J Cell Mol Med, 2020, 24(3):2123-2134] PubMed: 31957179
The Synergism of the Small Molecule ENOblock and Fluconazole Against Fluconazole-Resistant Candida albicans. [ Front Microbiol, 2019, 10:2071] PubMed: 31555252
Structural and Functional Studies of Bacterial Enolase, a Potential Target Against Gram-Negative Pathogens. [Krucinska J Biochemistry, 2018, 10.1021/acs.biochem.8b01298] PubMed: 30714720

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SHIPPING AND STORAGE
Selleck products are transported at room temperature. If you receive the product at room temperature, please rest assured, the Selleck Quality Inspection Department has conducted experiments to verify that the normal temperature placement of one month will not affect the biological activity of powder products. After collecting, please store the product according to the requirements described in the datasheet. Most Selleck products are stable under the recommended conditions.

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