Abstract 3394: Ponatinib, a pan-BCR-ABL inhibitor, potently inhibits key activating and drug-resistant KIT mutants found in GIST.

Ponatinib(AP24534) is a multi-targeted tyrosine kinase inhibitor with potent pan-BCR-ABL activity being investigated in patients with CML. Ponatinibalso inhibits the kinase activity of KIT, a clinically proven oncogenic driver. Approximately 80% of gastrointestinal stromal tumors( GIST) contain primary activating KIT mutations, the majority of which cluster in exon 11 (V560D, Δ557-8) or more rarely in exon 9 (AY502-3ins). Imatinib is approved for the 1 st line treatment of GIST; however, patients frequently relapse due to the acquisition of secondary resistance mutationslocated in either the KIT ATP-binding pocket (T670I, V654A) or the activation (A) loop (D816H, D820A, N822K, A829P). Sunitinibis registered for 2 nd line treatment of GISTbut does not effectively inhibit A-loop mutants. Here we explored the activity of ponatinibagainst major primary and secondary KIT mutants found in GIST. The drug sensitivity of KIT mutants was determined using engineered Ba/F3 cells harboring mutant forms of KIT. Imatinib potently inhibited the viability of cells expressing exon 11 mutant KIT (IC 50 50 180 nM - 10 μM). Sunitinibwas highly active (IC 50 50 > 200 nM). In contrast, ponatinibwas highly potent (IC 50 50 = 56 nM) or when V654A was present as a secondary mutation (IC 50 = 59 nM). Importantly, in patients dosed once daily with 45 mg ponatinib, peak (145 nM) and trough (64 nM) plasma concentrations achieved are predicted to lead to substantial inhibition of all KIT mutants tested here. To rationalize ponatinib9s activity we solved the structure of the native KIT kinase domain co-crystallized with ponatinib. Ponatinibbound the inactive (DFG-out) conformation of KIT, filling the ATP-binding pocket as well as two adjacent hydrophobic pockets, analogous to its binding to ABL. Ponatinibcan accommodate the gatekeeper T670I mutation by virtue of its triple bondcircumventing the steric bulk of the larger isoleucine residue. Ponatinib9s activity against A-loop mutants can be rationalized by its tight binding to the inactive conformation of KIT, even in the presence of A-loop mutations. In conclusion, ponatinibpossesses potent activity versus the major clinically relevant KIT mutants, inhibiting their activity with IC 50 s that fall within clinically achievable plasma concentrations. These data therefore support the evaluation of ponatinibin patients with GIST, particularly since KIT A-loop mutants represent an unmet medical need. Citation Format: Andrew P. Garner, Rana Anjum, Sadanand Vodala, Alexa Schrock, Tianjun Zhou, Tim Clackson, Joseph M. Gozgit, Victor M. Rivera. Ponatinib, a pan-BCR-ABL inhibitor, potently inhibits key activating and drug-resistant KIT mutants found in GIST. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3394. doi:10.1158/1538-7445.AM2013-3394