PH-797804 (PH797804), a diarylpyridinone analog, is a novel, potent, selective and ATP-competitive inhibitor of p38 mitogen-activated protein (p38α), with potential anti-inflammatory and anticancer activity. In a cell-free assay, it inhibits p38α with an IC50 of 26 nM, demonstrates a 4-fold preference for p38α over p38β, and has no effects on JNK2. Excellent anti-proliferative activity against colon tumor cells and high in vivo antitumor efficacy are both displayed by PH-797804 in three PDXs. It might be helpful in the treatment of colon cancer and rheumatoid arthritis.

Physicochemical Properties

Molecular Formula	C22H19BRF2N2O3
Molecular Weight	477.3
Exact Mass	476.054
Elemental Analysis	C, 55.36; H, 4.01; Br, 16.74; F, 7.96; N, 5.87; O, 10.06
CAS #	586379-66-0
Related CAS #	586379-66-0
PubChem CID	22049997
Appearance	White to off-white solid powder
Density	1.5±0.1 g/cm3
Boiling Point	593.2±50.0 °C at 760 mmHg
Flash Point	312.6±30.1 °C
Vapour Pressure	0.0±1.7 mmHg at 25°C
Index of Refraction	1.629
LogP	3.24
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	5
Rotatable Bond Count	5
Heavy Atom Count	30
Complexity	742
Defined Atom Stereocenter Count	0
SMILES	O=C(C1=CC=C(C(N2C(C)=CC(OCC3=CC=C(C=C3F)F)=C(C2=O)Br)=C1)C)NC
InChi Key	KCAJXIDMCNPGHZ-UHFFFAOYSA-N
InChi Code	InChI=1S/C22H19BrF2N2O3/c1-12-4-5-14(21(28)26-3)9-18(12)27-13(2)8-19(20(23)22(27)29)30-11-15-6-7-16(24)10-17(15)25/h4-10H,11H2,1-3H3,(H,26,28)
Chemical Name	3-[3-bromo-4-[(2,4-difluorophenyl)methoxy]-6-methyl-2-oxopyridin-1-yl]-N,4-dimethylbenzamide
Synonyms	PH 797804; PH797804; PH-797804
HS Tariff Code	2934.99.9001
Storage	Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month
Shipping Condition	Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Biological Activity

Targets	p38α (IC50 = 26 nM); p38α (Ki = 5.8 nM); p38β (Ki = 40 nM)
ln Vitro	PH-797804 blocks LPS-induced TNF-α production and p38 kinase activity in the human monocytic U937 cell line, with comparable IC50 of 5.9 nM and 1.1 nM. At concentrations up to 1 μM, PH-797804 has no inhibitory impact on the JNK pathway (c-Jun phosphorylation) or ERK pathway (ERK phosphorylation) in U937 cells. With an IC50 of 3 nM in primary rat bone marrow cells, PH-797804 inhibits RANKL- and M-CSF-induced osteoclast formation in a concentration-dependent manner.[1] The activity of PH-797804 is specific because the IC50 values for it against the following targets have been found to be higher than 200 μM (unless otherwise noted): CDK2, ERK2, IKK1, IKK2, IKKi, MAPKAP2, MAPKAP3, MKK7 (>100 μM), MNK, MSK (>164 μM), PRAK, RSK2, and TBK1.[2]
ln Vivo	In both rats and cynomolgus monkeys, oral administration of PH-797804 effectively reduces the acute inflammatory reactions brought on by systemically administered endotoxin. In chronic disease models, PH-797804 treatment for 10 days shows strong anti-inflammatory activity, significantly lowering joint inflammation and related bone loss in rats with streptococcal cell wall-induced arthritis and mice with collagen-induced arthritis. In rats and cynomolgus monkeys, the ED50 values were 0.07 mg/kg and 0.095 mg/kg, respectively, according to dose-response analysis. In a human endotoxin challenge model, PH-797804 inhibits LPS-induced TNF-α, IL-6, and MK-2 activity in a dose- and concentration-dependent manner.[1]
Enzyme Assay	The phosphorylation of GST-c-Jun or the epidermal growth factor receptor peptide (EGFRP) by p38 kinases is assessed using a resin capture assay method. 25 mM HEPES, pH 7.5, 10 mM magnesium acetate, 0.05 to 0.3 μCi of [γ-33P]ATP, 0.8 mM dithiothreitol, and either 200 μM EGFRP or 10 μM GST-c-Jun for p38 kinase reactions are included in reaction mixtures. To begin the reaction, 25 nM p38α kinase is added, resulting in a final volume of 50 μl. For 30 minutes, the p38α kinase reactions are incubated at 25 °C. Under these circumstances, both p38 kinase's product formation is time-dependently linear. The addition of 150 μl of AG 1 × 8 ion exchange resin in 900 mM sodium formate, pH 3.0, stops the reaction and removes the unreacted [γ-33P]ATP. Solutions are thoroughly combined and then left to stand for 5 minutes. The phosphorylated substrate is extracted from the mixture in a 50-μl aliquot, which is then transferred to a 96-well plate. A TopCount NXT microplate scintillation and luminescence counter is used to add 150 μL of the MicroScint-40 scintillation cocktail to each well and measure the radioactivity levels.
Cell Assay	The 3-(4,5-dimethylthiazol-2-yl)-) diphenyl tetrazolium bromide assay is used to measure cell viability. With a reference wavelength of 630 nm and a test wavelength of 570 nm, absorbance is measured on an ELISA plate reader.
Animal Protocol	LPS-induced chronic inflammation rat model 0.001-1 mg/kg Oral gavage 4 hours before LPS administration
References	[1]. Anti-inflammatory properties of a novel N-phenyl pyridinone inhibitor of p38 mitogen-activated protein kinase: preclinical-to-clinical translation. J Pharmacol Exp Ther, 2009, 331(3), 882-895. [2]. Structural bioinformatics-based prediction of exceptional selectivity of p38 MAP kinase inhibitor PH-797804. Biochemistry, 2009, 48(27), 6402-6411.
Additional Infomation	PH 797804 is a member of the class of benzamides obtained by formal condensation of the carboxy group of 3-{3-bromo-4-[(2,4-difluorobenzyl)oxy]-6-methyl-2-oxopyridin-1-yl}-4-methylbenzoic acid with the amino group of methylamine. It has a role as an EC 2.7.11.24 (mitogen-activated protein kinase) inhibitor and an anti-inflammatory agent. It is a member of benzamides, an organofluorine compound, a pyridone, an organobromine compound and an aromatic ether. PH-797804 has been investigated for the treatment of Osteoarthritis.

Solubility Data

Solubility (In Vitro)

DMSO: ~96 mg/mL (~201.1 mM) Water: <1 mg/mL Ethanol: ~7 mg/mL (~14.7mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 3 mg/mL (6.29 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 30.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (5.24 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. Solubility in Formulation 3: 2% DMSO+30% PEG 300+5% Tween 80+ddH2O: 5mg/mL  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.0951 mL	10.4756 mL	20.9512 mL
	5 mM	0.4190 mL	2.0951 mL	4.1902 mL
	10 mM	0.2095 mL	1.0476 mL	2.0951 mL

*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.