Inhibitors, Agonists, Screening Libraries
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Data Sheet
Product Name:Cat. No.:CAS No.:
Molecular Formula:Molecular Weight:Target:Pathway:Solubility:
GSK1379725AHY-1123981802251-00-8C23H23FN6O3450.47OthersOthersDMSO
BIOLOGICAL ACTIVITY:
GSK1379725A is an inhibitor of Brd4 and a selective BPTF ligand (Kd=2.8 μM).
In Vitro: From the NMR titration of GSK1379725A, the bound and unbound resonances are separated by 171 Hz, providing an upper
bound for the chemical exchange rate. Assuming an association rate of 1×108 M-1 s-1 as a high end for a range of protein-smallmolecule interactions, (e.g., chymotrypsin: proflavin k1=1.2×108 M-1 s-1), an upper Kd of 8 μM is estimated from this experiment. For amore accurate determination with a non-fluorinated protein, ITC is used as a complementary direct binding assay using unlabeledBPTF. A Kd of 2.8 μM is obtained, consistent with our intermediate exchange resonance broadening by PrOF NMR. Although
GSK1379725A has been demonstrated to be selective over Brd4, a full selectivity panel against other bromodomains will be needed. Adatabase search using ChEMBL only showed GSK1379725A to be active in five cellular assays with an EC50 of 500 nM carried out.Additionally, no kinase activity has been reported for GSK1379725A despite the growing screening use of the PKIS library[1].
PROTOCOL (Extracted from published papers and Only for reference)
Cell Assay:[1]Cell viability assay of HEK 293 cells are performed using resazurine dye based CellTiter-Blue. HEK 293T cells are plated in96 well plates and experiments conducted when cells are 80% confluent. Cells are treated with the 0, 1.0, 3.0 and 10.0 μMGSK1379725A (AU1) for 24 and 48 hours in 5% CO2 (n=5-6, per condition). 20 μL of CellTiter-Blue is added to the each well andincubated for 2.5 h at 37°C. As resazurin dye is reduced by viable cells to resorufin. Resorufin is fluorescent at 580 excitation and 590emission. The data is normalized with the control (DMSO treated)[1].
References:
[1]. Urick AK, et al. Dual Screening of BPTF and Brd4 Using Protein-Observed Fluorine NMR Uncovers New Bromodomain Probe Molecules. ACS Chem Biol.2015 Oct 16;10(10):2246-56.
Caution: Product has not been fully validated for medical applications. For research use only.
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