PD 173074 (Synonyms: PD 173074,PD-173074)

In vitro kinase inhibition assays

Assays using the full-length FGFR-1 kinase were performed in a total volume of 100 μL containing 25 mM HEPES buffer (pH 7.4), 150 mM NaCl, 10 mM MnCl2, 0.2 mM sodium orthovanadate, 750 μg/mL concentration of a random copolymer of glutamic acid and tyrosine (4:1), various concentrations of PD173074 and 60 to 75 ng of enzyme. The reaction was initiated by the addition of [γ-32P]ATP (5 μM ATP containing 0.4 μCi of [γ-32P]ATP per incubation), and samples were incubated at 25 °C for 10 mins. The reaction was terminated by the addition of 30% trichloroacetic acid and the precipitation of material onto glass-fiber filter mats. Filters were washed three times with 15% trichloroacetic acid, and the incorporation of [32P] into the glutamate tyrosine polymer substrate was determined by counting the radioactivity retained on the filters in a Wallac 1250 betaplate reader.

Cell lines

NIH 3T3 cells

Preparation method

The solubility of this compound in DMSO is >10 mM. General tips for obtaining a higher concentration: Please warm the tube at 37 °C for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below -20 °C for several months.

Reaction Conditions

0 ~ 1000 nM; 5 mins

Applications

PD 173074 dose-dependently inhibited autophosphorylation of FGFR1, with an IC50 value in the range of 1 ~ 5 nM. In addition, PD 173074 inhibited autophosphorylation of VEGFR2 with an IC50 value of 100 ~ 200 nM.

Animal models

Swiss Webster mice with induced corneal angiogenesis

Dosage form

1 or 2 mg/kg/day; i.p.

Applications

At the dose of 1 or 2 mg/kg, PD 173074 significantly inhibited angiogenesis induced by either FGF or VEGF in a dose-dependent manner. Besides, it showed no apparent toxicity.

Other notes

Please test the solubility of all compounds indoor, and the actual solubility may slightly differ with the theoretical value. This is caused by an experimental system error and it is normal.

References:

[1]. Mohammadi M, Froum S, Hamby JM, Schroeder MC, Panek RL, Lu GH, Eliseenkova AV, Green D, Schlessinger J, Hubbard SR. Crystal structure of an angiogenesis inhibitor bound to the FGF receptor tyrosine kinase domain. EMBO J. 1998 Oct 15;17(20):5896-904.