Molecule

Description

Glioma, Cancer

Description

Crenolanib (CP-868569) is a potent and selective inihibitor of PDGFRα with IC50 of 10 nM in PDGFRα over-expressed CHO cells.

Targets

IC₅₀

In Vitro

Crenolanib (CP-868569) is significantly more potent than imatinib in inhibiting the kinase activity of imatinib-resistant PDGFRα kinases (D842I, D842V, D842Y, D1842-843IM, and deletion I843). Crenolanib (CP-868569) is 135-fold more otent than imatinib against D842V in the isogenic model system, with an IC50 of approximately 10 nM. Crenolanib (CP-868569) inhibits the kinase activity of the fusion oncogene in EOL-1 cell line, which is derived from a patient with chronic eosinophilic leukemia and expresses the constitutively activated FIP1L1- PDGFRα fusion kinase, with IC50 = 21 nM. Crenolanib (CP-868569) also inhibits the proliferation of EOL-1 cells with IC50 = 0.2 pM. Crenolanib (CP-868569) inhibits the activation of V561D or D842V –mutant kinases expressed in BaF3 cells with IC50 with 85 nM or 272 nM, respectively. Crenolanib (CP-868569) inhibits PDGFRα activation in H1703 non-small cell lung cancer cell line which has 24-fold amplification of the 4q12 region that contains the PDGFRα locus, with IC50 with 26 nM. ^[1] CP-868,596 is an orally bioavailable, highly potent and selective PDGFR TKI. CP-868,596 is a benzimidazole compound that has IC50s of 0.9 nM and 1.8 nM for PDGFRA and PDGFRB, respectively. ^[2]

In Vivo

Clinical Trials

Features

Biochemical Assessment of PDGFRα Kinase Activity

Chinese hamster ovary (CHO) cells are transiently transfected with mutated or wild type PDGFRα constructs and treated with various concentrations of Crenolanib (CP-868569). Experiments involving recombinant DNA are performed using biosafety level 2 conditions in accordance with guidelines. Protein lysates from cell lines are prepared and subjected to immunoprecipitation using anti-PDGFRα antibodies followed by sequential immunoblotting for PDGFRα. Densitometry is performed to quantify drug effect using Photoshop software, with the level of phosphor- PDGFRα normalized to total protein. Densitometry and proliferation experimental results are analyzed using Calcusyn 2.1 software to mathematically determine the IC50 values. The Wilcoxon Rank Sum Test is used to compare the IC50 values of Crenolanib (CP-868569) for a given mutation.

Cell Lines

EOL-1 cell line

Concentrations

0 – 20 pM

Incubation Time

72 hours

Methods

Cells are added to 96-well plates at densities of 20, 000 cells/well and incubated with Crenolanib (CP-868569) for 72 hours before measuring cellular proliferation using a 2,3-bis[2-methoxyl-4-nitro-5-sulfophenyl]-2H-tetrazolium-5-carboxanilide (XTT)-based assay.