Inhibitors, Agonists, Screening Libraries
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Data Sheet
Product Name:Cat. No.:CAS No.:
Molecular Formula:Molecular Weight:Target:Pathway:Solubility:
SB–505124HY-13521694433-59-5C20H21N3O2335.40
TGF–β ReceptorTGF–beta/Smad10 mM in DMSO
BIOLOGICAL ACTIVITY:
SB–505124 is a selective inhibitor of TGFβR with IC50 of 129 nM and 47 nM for ALK4, ALK5, respectively, and it does not inhibits ALK1,2, 3, or 6 but ALK7.
IC50 & Target: IC50: 129 nM (ALK4), 47 nM (ALK5)
In Vitro: SB–505124 demonstrates no toxicity to renal epithelial A498 cells at concentrations up to 100 μM for 48 h. 505124 inhibits
the closely related ALK4 with an IC50 value of 129±11 nM (about 2.5–fold less sensitive than ALK5) but does not inhibit ALK2 atconcentrations up to 10 μM. SB–505124 (1 μM) inhibits the TGF–β–induced phosphorylation of Smad2 in all three of these cell linesin a concentration–dependent fashion. SB–505124 (1 or 5 μM) potently inhibits TGF–β–induced activation of JNK/SAP, extracellularphosphorylation and CTGF and α–SMA expression in vitro[2]. SB–505124 susspresses CTGF and α–SMA observed byoutgrowth is poor from those treated with MMC[3].
signal–regulated kinase 1/2, and p38 despite the different patterns of activation in these cells[1]. SB–505124 (10 μM) impairs Smad2immunofluorescence. Cell outgrowth from explants dissected from eyes to which SB–505124 is applied during GFS is robust while
PROTOCOL (Extracted from published papers and Only for reference)
Kinase Assay:[1]Briefly, phospho–p38 is immunoprecipitated from 200 μg of cell lysates with an immobilized phosphor–p38 antibodyovernight at 4°C. p38 kinase activity is measured using 2 μg of ATF–2 fusion protein as the substrate with addition of 200 μM ATP.After a 30–min incubation at 30°C, the reaction is terminated with LaemmLi sample buffer, and the proteins are boiled and resolved by10% SDS–polyacrylamide gel electrophoresis, transferred to nitrocellulose membrane, and immunoblotted with phospho–ATF–2antibody.
Cell Assay:[1]Cell viability is measured as described previously or by using the modified tetrazolium salt WST–1. Approximately 2000cells are seeded in 96–well dishes in 100 μL of 0.2% FBS phenol red–free media overnight. The cells are treated with 50 μL of
SB–505124 (to achieve the final concentrations indicated) for 30 min before being treated with or without TGF–β1 and TNF–α to a finalvolume of 200 μL. Cell growth is measured at the indicated time points by incubating each well with 10 μL of WST–1 for 3 h at 37°C.Metabolically active cells cleave WST–1 to water–soluble formazan, which is directly quantitated with an enzyme–linkedimmunosorbent assay plate reader. Each experiment is done at least twice, and treatment for each cell line is done in triplicate.
References:
[1]. DaCosta Byfield S, et al. SB–505124 is a selective inhibitor of transforming growth factor–beta type I receptors ALK4, ALK5, and ALK7. Mol Pharmacol.2004 Mar;65(3):744–52.
[2]. Sutariya V, et al. Thermoreversible gel for delivery of activin receptor–like kinase 5 inhibitor SB–505124 for glaucoma filtration surgery. Pharm DevTechnol. 2013 Jul–Aug;18(4):957–62.
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[3]. Sapitro J, et al. Suppression of transforming growth factor–β effects in rabbit subconjunctival fibroblasts by activin receptor–like kinase 5 inhibitor. MolVis. 2010 Sep 16;16:1880–92.
Caution: Product has not been fully validated for medical applications. For research use only.
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