PH-797804

LPS-induced chronic inflammation rat model

3-(4-(2, 4-difluorobenzyloxy)-3-bromo-6-methyl-2-oxopyridin-1(2H)-yl)-N, 4-dimethylbenzamide

PH-797804 is a novel pyridinone inhibitor of p38alpha with IC50 of 26 nM.

PH-797804 is prepared as a suspension in a vehicle consisting of 0.5% methylcellulose and 0.025% Tween 20.

PH-797804 blocks LPS-induced TNF-alpha production and p38 kinase activity in the human monocytic U937 cell line, with comparable IC50 of 5.9 nM and 1.1 nM. PH-797804 has no inhibitory effect on either the JNK pathway (c-Jun phosphorylation) or ERK pathway (ERK phosphorylation) in U937 cells at concentrations up to 1 uM. PH-797804 inhibits RANKL- and M-CSF-induced osteoclast formation in a concentration-dependent manner, with IC50 of 3 nM in primary rat bone marrow cells. [1] IC50 values for PH-797804 against the following targets have been determined to be greater than 200 uM (unless specified): CDK2, ERK2, IKK1, IKK2, IKKi, MAPKAP2, MAPKAP3, MKK7 (>100 uM), MNK, MSK (>164 uM), PRAK, RSK2, and TBK1, which means the activity of PH-797804 is specific. [2]

P38 kinase assay, A resin capture assay method is used to determine the phosphorylation of epidermal growth factor receptor peptide (EGFRP) or GST-c-Jun by p38 kinases. Reactions mixtures contain 25 mM HEPES, pH 7.5, 10 mM magnesium acetate, ATP (at the indicated concentration), 0.05 to 0.3 uCi of [gamma-33P]ATP, 0.8 mM dithiothreitol, and either 200 uM EGFRP or 10 uM GST-c-Jun for p38alpha kinase reactions. The reaction is initiated by the addition of 25 nM p38alpha kinase to give a final volume of 50 ul. The p38alphakinase reactions are incubated at 25 C for 30 minutes. Under these conditions, the formation of product for both p38alphakinase is linear with time. The reaction is stopped, and the unreacted [gamma-33P]ATP is removed by the addition of 150 ul of AG 1, 8 ion exchange resin in 900 mM sodium formate, pH 3.0. Once thoroughly mixed, solutions are allowed to stand for 5 minutes. A 50-ul aliquot of head volume containing the phosphorylated substrate is removed from the mixture and transferred to a 96-well plate. MicroScint-40 scintillation cocktail (150 uL) is added to each well and the radioactivity quantities using a TopCount NXT microplate scintillation and luminescence counter.

477, 3

DMSO 96 mg/mL, Water <1 mg/mL, Ethanol 7 mg/mL