Resistance to a tyrosine kinase inhibitor mediated by changes to the conformation space of the kinase

• Published in: Physical chemistry chemical physics : PCCP 2023-02, Vol.25 (8), p.6175-6183
• Main Authors: Freire, Thales Souza, Caracelli, Ignez, Zukerman-Schpector, Julio, Friedman, Ran
• Format: Article
• Language: English
• Subjects: Aniline Compounds; Biochemistry; Biokemi; Cell Line, Tumor; Enzymes; fms-Like Tyrosine Kinase 3; Gibbs free energy; Kinases; Leukemia; Mathematical analysis; Mutation; Protein Kinase Inhibitors - pharmacology; Proteins; Tyrosine; Tyrosine Kinase Inhibitors
• ISSN: 1463-9076; 1463-9084; 1463-9084
• DOI: 10.1039/d2cp05549j

Gilteritinib is a highly selective and effective inhibitor of the FLT3/ITD mutated protein, and is used successfully in treating acute myeloid leukaemia (AML). Unfortunately, tumour cells gradually develop resistance to gilteritinib due to mutations in the molecular drug target. The atomistic details behind this observed resistance are not clear, since the protein structure of the complex is only available in the inactive state, while the drug binds better to the active state. To overcome this limitation, we used a computer-aided approach where we docked gilteritinib to the active site of FLT3/ITD and calculated the Gibbs free energy difference between the binding energies of the parental and mutant enzymes. These calculations agreed with experimental estimations for one mutation (F691L) but not the other (D698N). To further understand how these mutations operate, we used metadynamics simulations to study the conformational landscape of the activation process. Both mutants show a lower activation energy barrier which suggests that they are more likely to adopt an active state until inhibited, making the mutant enzymes more active. This suggests that a higher efficiency of tyrosine kinases contributes to resistance not only against type 2 but also against type 1 kinase inhibitors. Gilteritinib is an effective inhibitor of the FLT3/ITD mutated protein, and is used in treating acute myeloid leukaemia (AML), but resistance due to mutations in the protein is a problem. We studied how resistance is mediated using advanced simulation methods.