IOX1

Catalog No.S7234 Batch:S723403

Technical Data

Formula	C₁₀H₇NO₃
Molecular Weight	189.17	CAS No.	5852-78-8
Solubility (25°C)*	In vitro	DMSO	38 mg/mL (200.87 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.)

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Biological Activity

Description

IOX1 is a potent and broad-spectrum inhibitor of 2OG oxygenases, including the JmjC demethylases. IOX1 is an inhibitor of ALKBH5.

Targets

KDM3A ^[1] (Cell-free assay)	KDM4C ^[1] (Cell-free assay)	KDM6B ^[1] (Cell-free assay)	KDM2A ^[1] (Cell-free assay)	KDM4E ^[1] (Cell-free assay)	View More
0.1 μM	0.6 μM	1.6 μM	1.8 μM	2.3 μM	View More

In vitro

IOX1 increases H3K9me3 levels in HeLa cells via KDM4A inhibition without significant effect on cell viability. IOX1 shows lower efficacy in HeLa cells due to low cell permeability, while the n-octyl ester derivative improves its cell permeability. ^[1]

Features

Cell-permeant, broad-spectrum 2OG oxygenase inhibitor.

Protocol (from reference)

Kinase Assay:^{^[1]}	AlphaScreen Assay All reagents are diluted in 50 mM HEPES, 0.1% BSA, pH 7.5 supplemented with 0.01% Tween20 and allowed to equilibrate to room temperature prior to addition to plates. Catalytic turnover assays are run in 10 μL volumes in lowvolume 384-well plates at RT. The reaction consisted of enzyme (5 nM), biotinylated substrate peptide (30 nM), Fe(II) (1 μM), ascorbate (100 μM), 2OG (10 μM) and run at RT. For PHD2, the reaction consisted of enzyme (5 nM), biotinylated substrate peptide (60nM), Fe(II) (20 μM), ascorbate (200 μM), 2OG (2 μM) and run at RT. EDTA is used to quench the reaction (5 μL), AlphaScreen donor (Streptavidin-conjugated) and acceptor (Protein A-conjugated) beads preincubated with peptide product antibodies are added (5 μL). Plates are foil-sealed to protect from light, incubated at room temperature for 60 minutes and read on a PHERAstar FS plate reader using an AlphaScreen 680 excitation/570 emission filter set. The final bead concentration in 20 μL reaction is 20 μg/mL. IC50 values are calculated in Prism 6 after normalisation against corresponding DMSO controls.
Cell Assay:^{^[1]}	Cell lines HeLa cells Concentrations ~300 μ M Incubation Time 24 hours Method Antiproliferative activities of compounds are determined by the MTT assay. HeLa cells are seeded into 96-well plates and cultured at 37 °C for 24 h to achieve 70%. Subsequently, the medium is replaced with DMEM medium containing the tested compounds in different concentrations of 1-300 μM in 1% DMSO. Staurosporine in 0.03-10 μM final concentration is used as a control for cytotoxicity. After 24 h of treatment, the medium is replaced with CellTiter 96 Aqueous One Solution Reagent and incubated for 4 hours. CC50 values are calculated in Prism 6 software after normalisation against corresponding 1% DMSO treated cells and 1% DMSO in media (no cells) controls.

Kinase Assay:^{^[1]}

AlphaScreen Assay
All reagents are diluted in 50 mM HEPES, 0.1% BSA, pH 7.5 supplemented with 0.01% Tween20 and allowed to equilibrate to room temperature prior to addition to plates. Catalytic turnover assays are run in 10 μL volumes in lowvolume 384-well plates at RT. The reaction consisted of enzyme (5 nM), biotinylated substrate peptide (30 nM), Fe(II) (1 μM), ascorbate (100 μM), 2OG (10 μM) and run at RT. For PHD2, the reaction consisted of enzyme (5 nM), biotinylated substrate peptide (60nM), Fe(II) (20 μM), ascorbate (200 μM), 2OG (2 μM) and run at RT. EDTA is used to quench the reaction (5 μL), AlphaScreen donor (Streptavidin-conjugated) and acceptor (Protein A-conjugated) beads preincubated with peptide product antibodies are added (5 μL). Plates are foil-sealed to protect from light, incubated at room temperature for 60 minutes and read on a PHERAstar FS plate reader using an AlphaScreen 680 excitation/570 emission filter set. The final bead concentration in 20 μL reaction is 20 μg/mL. IC50 values are calculated in Prism 6 after normalisation against corresponding DMSO controls.

Cell Assay:^{^[1]}

Cell lines
HeLa cells
Concentrations
~300 μ M
Incubation Time
24 hours
Method
Antiproliferative activities of compounds are determined by the MTT assay. HeLa cells are seeded into 96-well plates and cultured at 37 °C for 24 h to achieve 70%. Subsequently, the medium is replaced with DMEM medium containing the tested compounds in different concentrations of 1-300 μM in 1% DMSO. Staurosporine in 0.03-10 μM final concentration is used as a control for cytotoxicity. After 24 h of treatment, the medium is replaced with CellTiter 96 Aqueous One Solution Reagent and incubated for 4 hours. CC50 values are calculated in Prism 6 software after normalisation against corresponding 1% DMSO treated cells and 1% DMSO in media (no cells) controls.

References

Selleck's IOX1 has been cited by 12 publications

Characterization of the m6A regulators' landscape highlights the clinical significance of acute myocardial infarction [ Front Immunol, 2024, 15:1308978]	PubMed: 38571952
Inhibition of ALKBH5 attenuates I/R-induced renal injury in male mice by promoting Ccl28 m6A modification and increasing Treg recruitment [ Nat Commun, 2023, 14(1):1161]	PubMed: 36859428
Oxygen-sensitive methylation of ULK1 is required for hypoxia-induced autophagy [ Nat Commun, 2022, 13(1):1172]	PubMed: 35246531
Discovery of two novel ALKBH5 selective inhibitors that exhibit uncompetitive or competitive type and suppress the growth activity of glioblastoma multiforme [ Chem Biol Drug Des, 2022, 10.1111/cbdd.14051]	PubMed: 35384315
IOX1 activity as sepsis therapy and an antibiotic against multidrug-resistant bacteria [ Sci Rep, 2021, 11(1):2942]	PubMed: 33536477
IOX1 impedes host inflammation in imiquimod-triggered psoriasis [ Heliyon, 2021, 7(11):e08433]	PubMed: 34877426
KDM3A Senses Oxygen Availability to Regulate PGC-1α-Mediated Mitochondrial Biogenesis. [ Mol Cell, 2019, 76(6):885-895]	PubMed: 31629659
The histone demethylase JMJD2B is critical for p53-mediated autophagy and survival in Nutlin-treated cancer cells [ J Biol Chem, 2019, 294(23):9186-9197]	PubMed: 31036564
Epigenetic Reprogramming with Antisense Oligonucleotides Enhances the Effectiveness of Androgen Receptor Inhibition in Castration-Resistant Prostate Cancer [ Cancer Res, 2018, 78(20):5731-5740]	PubMed: 30135193
[ Cell Death Dis, 2018, ]	PubMed: 29988031

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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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