Imatinib, which is an inhibitor of the
BCR-ABL tyrosine kinase, has been a remarkable success for the treatment of
Philadelphia chromosome-positive (Ph+)
chronic myelogenous leukemias (CMLs). However, a significant proportion of patients chronically treated with
imatinib develop resistance because of the acquisition of mutations in the
kinase domain of BCR-ABL. Mutations occur at residues directly implicated in
imatinib binding or, more commonly, at residues important for the ability of the
kinase to adopt the specific closed (inactive) conformation to which
imatinib binds. In our quest to develop new BCR-ABL inhibitors, we chose to target regions outside the
ATP-binding site of this
enzyme because these compounds offer the potential to be unaffected by mutations that make CML cells resistant to
imatinib. Here we describe the activity of one compound,
ON012380, that can specifically inhibit BCR-ABL and induce cell death of Ph+ CML cells at a concentration of <10 nM. Kinetic studies demonstrate that this compound is not
ATP-competitive but is substrate-competitive and works synergistically with
imatinib in wild-type BCR-ABL inhibition. More importantly,
ON012380 was found to induce apoptosis of all of the known
imatinib-resistant mutants at concentrations of <10 nM concentration in vitro and cause regression of
leukemias induced by i.v. injection of 32Dcl3 cells expressing the
imatinib-resistant BCR-ABL
isoform T315I. Daily i.v. dosing for up to 3 weeks with a >100 mg/kg concentration of this agent is well tolerated in rodents, without any hematotoxicity.