HS-243

Catalog No.S9701 Batch:S970101

Technical Data

Formula	C₁₇H₁₆N₄O₃
Molecular Weight	324.33	CAS No.	848249-10-5
Solubility (25°C)*	In vitro	DMSO	16 mg/mL (49.33 mM)
		Water	Insoluble
		Ethanol	Insoluble
* <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

HS-243 is a highly potent interleukin-1 receptor-associated kinase (IRAK) inhibitor with IC50 of 24 nM, 20 nM and 0.5 μM for IRAK-1, IRAK-4 and TAK1, respectively.

Targets

IRAK-4 ^[1] (Cell-free assay)	IRAK-1 ^[1] (Cell-free assay)	TAK1 ^[1] (Cell-free assay)
20 nM	24 nM	0.5 μM

In vitro

HS-243 has exquisite selectivity toward both IRAK-1 (IC50 = 24 nM) and IRAK-4 (IC50 = 20 nM), with only minimal TAK1-inhibiting activity (IC50 = 0.5 μM). Using HS-243 and takinib, we evaluates the consequences of cytokine/chemokine responses after selective inhibition of IRAK-1/4 or TAK1 in response to lipopolysaccharide challenge in human rheumatoid arthritis fibroblast-like synoviocytes. HS-243 specifically inhibits intracellular IRAKs without TAK1 inhibition and these kinases have distinct, nonredundant signaling roles.^[1]

Protocol (from reference)

Kinase Assay:^{^[1]}	Activity of purified IRAK-4 protein In brief, IRAK-4 (20 ng/well) is incubated with 2 mM ATP containing radiolabeled [32P]ATP in the presence of 300 μM substrate peptide in a final volume of 40 μl in the presence of buffer and HS-243. The reaction is terminated at 20 min with 10 μl of 1 M H3PO4. The remaining activity is measured using a scintillation counter. Dose–response curves are repeated two times and averaged.
Cell Assay:^{^[1]}	Cell lines RA-FLS cells, THP-1 cells Concentrations 10 μM Incubation Time 24 h Method RA-FLS cells or THP-1 cells are treated with 10 μM HS-243 or vehicle (DMSO), and 24 h after treatment, supernatant is added to human cytokine XL proteome array in accordance with the manufacturer’s protocol. Chemiluminescence is used to visualize protein quantities.

Kinase Assay:^{^[1]}

Activity of purified IRAK-4 protein
In brief, IRAK-4 (20 ng/well) is incubated with 2 mM ATP containing radiolabeled [32P]ATP in the presence of 300 μM substrate peptide in a final volume of 40 μl in the presence of buffer and HS-243. The reaction is terminated at 20 min with 10 μl of 1 M H3PO4. The remaining activity is measured using a scintillation counter. Dose–response curves are repeated two times and averaged.

Cell Assay:^{^[1]}

Cell lines
RA-FLS cells, THP-1 cells
Concentrations
10 μM
Incubation Time
24 h
Method
RA-FLS cells or THP-1 cells are treated with 10 μM HS-243 or vehicle (DMSO), and 24 h after treatment, supernatant is added to human cytokine XL proteome array in accordance with the manufacturer’s protocol. Chemiluminescence is used to visualize protein quantities.

References

[1] Scott A Scarneo, et al. J Biol Chem. 2020 Feb 7;295(6):1565-1574.

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SHIPPING AND STORAGE
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