AP-III-a4 (ENOblock)

Catalog No.S7443 Batch:S744302

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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)
Ethanol 100 mg/mL (158.43 mM)
Water 9 mg/mL (14.25 mM)
In vivo (Add solvents to the product individually and in order)
Homogeneous suspension
CMC-NA
≥5mg/ml Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Clear solution
5%DMSO 40%PEG300 5%Tween80 50%ddH2O

Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.

 5.000mg/ml (7.92mM) Taking the 1 mL working solution as an example, add 50 μL of 100 mg/ml clarified DMSO stock solution to 400 μL of PEG300, mix evenly to clarify it; add 50 μL of Tween80 to the above system, mix evenly to clarify; then continue to add 500 μL of ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results. 
Clear solution
5% DMSO 95% Corn oil

Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.

 5.000mg/ml (7.92mM) Taking the 1 mL working solution as an example, add 50 μL of 100 mg/ml clear DMSO stock solution to 950 μL of corn oil and mix evenly. The mixed solution should be used immediately for optimal results. 
* <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 (ENOblock) 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, it also induces glucose uptake and inhibits phosphoenolpyruvate carboxykinase (PEPCK) expression.
In vivo In a HCT116-xenotransplanted zebrafish tumor xenograft model, AP-III-a4 (ENOblock) (10 μM) inhibits cancer cell migration and invasion processes. In vivo, it also causes downregulation of PEPCK expression and induction of glucose uptake, and inhibits adipogenesis and foam cell formation.

Protocol (from reference)

Kinase Assay:

[1]
  • Enolase activity assay

    Enolase activity 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 AP-III-a4 (ENOblock) or NaF. 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. 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

    AP-III-a4 (ENOblock) is used to treat 3 x 105 HCT116 cells seeded in a 6 well plate for 24 h, with or without prior induction of hypoxia for 4 h, using triplicate wells. After 24 h of treatment with this compound, cells are 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

    --

References

  • https://pubmed.ncbi.nlm.nih.gov/23547795/

Selleck's AP-III-a4 (ENOblock) Has Been Cited by 15 Publications

Targeting Kupffer Cell Enolase 1 Attenuates Liver Inflammation and Injury in Hemorrhagic Shock [ International Journal of Molecular Sciences, 2025, 8340] PubMed: 40943261.0
LncPRESS1 promotes the progression and cisplatin resistance of breast cancer through upregulation of ENO1 and activation of glycolysis [ Cell Division, 2025, 23] PubMed: 41046264.0
LncPRESS1 promotes the progression and cisplatin resistance of breast cancer through upregulation of ENO1 and activation of glycolysis [ Cell Div, 2025, 20(1):23] PubMed: 41046264
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
Mediation of PKM2-dependent glycolytic and non-glycolytic pathways by ENO2 in head and neck cancer development [ Journal of Experimental & Clinical Cancer Research, 2023, 1] PubMed: 36588153.0
Evolutionary genomics identifies host-directed therapeutics to treat intracellular bacterial infections [ bioRxiv, 2023, 10.1101/2023.08.01.551011] PubMed: None
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 [ Frontiers in Pharmacology, 2022, 807742] PubMed: 35095517.0
Amoxicillin Haptenation of α-Enolase is Modulated by Active Site Occupancy and Acetylation [ Front Pharmacol, 2021, 12:807742] PubMed: 35095517

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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.

NOT FOR HUMAN, VETERINARY DIAGNOSTIC OR THERAPEUTIC USE.