Cell experiment:

For cell proliferation assays, 45 μL containing 1000 cells per well are seeded into solid white 384-well plates in appropriate media. The following day, Dubermatinib (TP-0903) is diluted in serum free growth media to 10x desired concentrations and 5 μL is added to each well. Combined compound and cells are incubated for 96 hours. Following incubation, 40 μL of ATP-Lite solution is added to each well, incubated for an additional 10 minutes at room temperature and luminescence is measured on an microplate reader[1].

Description:

IC50: 0.027 μM against AXL

AXL and other TAM family members are known to be involved in maintaining the mesenchymal phenotype in cancer cells. Mesenchymal cells have increased invasion and migratory properties, enhanced cell survival in stressed environments, as well as increased resistance to targeted therapies. TP-0903 is a potent anti-cancer agent targeting the AXL receptor tyrosine kinase.

In vitro: On the basis of the potency of TP-0903 in biochemical assays, its activity in cell-based studies was evaluated. TP-0903 showed extremely potent activity in cell viability assays against the pancreatic cancer cell line PSN-1. More importantly, TP-0903 was evaluated for its ability to block GAS6-mediated activation of AXL in pancreatic cancer cells. PSN-1 cells were serum-starved and then stimulated with GAS6 in the presence of various concentrations of TP-0903 [1].

In vivo: TP-0903 restores sensitivity to erlotinib in cell-based and preclinical animal models of cancer by reversing the mesenchymal phenotype driving resistance [2].

Clinical trial: Up to now, TP-0903 is still in the preclinical development stage.

Reference:
[1] Mollard A, Warner SL, Call LT, Wade ML, Bearss JJ, Verma A, Sharma S, Vankayalapati H, Bearss DJ.  Design, Synthesis and Biological Evaluation of a Series of Novel Axl Kinase Inhibitors. ACS Med Chem Lett. 2011 Dec 8;2(12):907-912.
[2] ToleroPharmaceuticals, Inc–TP-0903.  http://www.toleropharmaceuticals.com/TP-0903.html.