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|English to Polish translations [PRO]|
Science - Biology (-tech,-chem,micro-)
|English term or phrase: recombinant kinase domain of the ALK|
|The efficacy of the compounds of formula (I) as inhibitors of ALK tyrosine kinase activity can be demonstrated as follows: The inhibition of ALK tyrosine kinase activity can be demonstrated using known methods, e.g., using the recombinant kinase domain of the ALK in analogy to the VEGF-R kinase assay. See Wood et al., Cancer Res, Vol. 60, No. 8, pp. 2178-2189 (2000). In vitro enzyme assays using GST-ALK protein tyrosine kinase are performed in 96-well plates as a filter binding assay in 20 mM tris·HCl, pH=7.5, 3 mM MgCl2, 10 mM MnCl2, 1 mM DTT, 0.1 PCi/assay (=30|
PL) [g-33P]-ATP, 2 PM ATP, 3 Pg/mL poly (Glu, Tyr 4:1) Poly-EY (Sigma P-0275), 1% DMSO, 25 ng ALK enzyme. Assays are incubated for 10 minutes at ambient temperature. Reactions are terminated by adding 50 PL of 125 mM EDTA, and the reaction mixture is transferred onto a MAIP Multiscreen plate (Millipore, Bedford, MA, USA), previously wet with methanol, and re-hydrated for 5 minutes with H2O. Following washing (0.5% H3PO4), plates are counted in a
liquid scintillation counter. IC50 values are calculated by linear regression analysis of the percentage inhibition. Compared with the control without inhibitor, the compounds of formula (I) inhibit the enzyme activity by 50% (IC50), e.g., in a concentration of from 0.001-0.5 PM, especially from 0.1-0.8 PM.
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