CH5183284 (Debio-1347)

Protein kinase assay

The inhibitory activity of CH5183284/Debio 1347 against FGFR1 was evaluated using a radiometric filter assay by measuring the incorporation of 33Pi with a microplate scintillation counter. The phosphorylation activities of LCK, EGFR, KIT, MET, SRC, BRK, FGFR2, Flt3, LTK, INSR, YES, ABL, EPHA2, ZAP70, Fyn, IGF1R, KDR and PDGFR on substrate peptides were determined by homogeneous time-resolved fluorescence assay with LANCE Eu-W1024 labeled anti-phosphotyrosine PT66 antibody according to standard methods. Time-resolved fluorescence was measured with an EnVision HTS microplate reader. The activities of Aurora A, Akt1/PKBα, PKA, Cdk1/cyclin B, Cdk2/cyclin A, PKCα, PKCβ1 and PKCβ2 on substrate peptides were determined by IMAP FP Screening Express Progressive Binding System. Fluorescence polarization was measured with an EnVision HTS microplate reader.

Cell lines

327 human tumor cell lines

Preparation method

The solubility of this compound in DMSO is > 17.8 mg/mL. 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.

Reacting condition

0.076 ~ 10,000 nM; 4 days

Applications

In DMS114 (FGFR1 amplification), SNU-16 (FGFR2 amplification) and KMS11 [t(4;14) translocation and FGFR3 Y373C mutation] cells, CH5183284 inhibited autophosphorylation of FGFR1, FGFR2 and FGFR3 at the dose range of 100 ~ 300 nM. Thus, CH5183284 selectively inhibited proliferation of cancer cell lines with genetic alterations in FGFR. In addition, CH5183284 also inhibited FGFR2-harboring cancer cells with V564F mutation which are resistant to other FGFR inhibitors.

Animal models

Mice bearing KG1, MFE280, SNU-16, RT112/84 or UM-UC-14 xenografts

Dosage form

100 mg/kg/day; p.o.

Applications

CH5183284 exhibited selective and significant antitumor activities against various xenografts with FGFR genetic alterations such as KG1 (leukemia, FGFR1OP-FGFR1 fusion), MFE-280 (endometrial cancer, FGFR2 S252W mutation), SNU-16 (gastric cancer, FGFR2 amplification), RT112/84 (bladder cancer, FGFR3-TACC3 fusion) and UM-UC-14 (bladder cancer, FGFR3 S249C mutation).

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]. Nakanishi Y, Akiyama N, Tsukaguchi T, et al. The fibroblast growth factor receptor genetic status as a potential predictor of the sensitivity to CH5183284/Debio 1347, a novel selective FGFR inhibitor. Mol Cancer Ther, 2014, 13(11): 2547-2558.