title: A drug resistance screen using a selective MET inhibitor reveals a spectrum of mutations that partially overlap with activating mutations found in cancer patients.
abstract: The emergence of drug resistance is a primary concern in any cancer treatment, including with targeted kinase inhibitors as exemplified by the appearance of Bcr-Abl point mutations in chronic myeloid leukemia (CML) patients treated with imatinib. In vitro approaches to identify resistance mutations in Bcr-Abl have yielded mutation spectra that faithfully recapitulated clinical observations. To predict resistance mutations in the receptor tyrosine kinase MET that could emerge during inhibitor treatment in patients, we conducted a resistance screen in BaF3 TPR-MET cells using the novel selective MET inhibitor NVP-BVU972. The observed spectrum of mutations in resistant cells was dominated by substitutions of tyrosine 1230 but also included other missense mutations and partially overlapped with activating MET mutations that were previously described in cancer patients. Cocrystallization of the MET kinase domain in complex with NVP-BVU972 revealed a key role for Y1230 in binding of NVP-BVU972, as previously reported for multiple other selective MET inhibitors. A second resistance screen in the same format with the MET inhibitor AMG 458 yielded a distinct spectrum of mutations rich in F1200 alterations, which is consistent with a different predicted binding mode. Our findings suggest that amino acid substitutions in the MET kinase domain of cancer patients need to be carefully monitored before and during treatment with MET inhibitors, as resistance may preexist or emerge. Compounds binding in the same manner as NVP-BVU972 might be particularly susceptible to the development of resistance through mutations in Y1230, a condition that may be addressed by MET inhibitors with alternative binding modes.
authors.affiliation: Novartis Institutes for BioMedical Research, Basel, Switzerland.
authors.forename: Ralph
authors.initials: R
authors.lastname: Tiedt
authors.forename: Elisa
authors.initials: E
authors.lastname: Degenkolbe
authors.forename: Pascal
authors.initials: P
authors.lastname: Furet
authors.forename: Brent A
authors.initials: BA
authors.lastname: Appleton
authors.forename: Sabrina
authors.initials: S
authors.lastname: Wagner
authors.forename: Joseph
authors.initials: J
authors.lastname: Schoepfer
authors.forename: Emily
authors.initials: E
authors.lastname: Buck
authors.forename: David A
authors.initials: DA
authors.lastname: Ruddy
authors.forename: John E
authors.initials: JE
authors.lastname: Monahan
authors.forename: Michael D
authors.initials: MD
authors.lastname: Jones
authors.forename: Jutta
authors.initials: J
authors.lastname: Blank
authors.forename: Dorothea
authors.initials: D
authors.lastname: Haasen
authors.forename: Peter
authors.initials: P
authors.lastname: Drueckes
authors.forename: Markus
authors.initials: M
authors.lastname: Wartmann
authors.forename: Clive
authors.initials: C
authors.lastname: McCarthy
authors.forename: William R
authors.initials: WR
authors.lastname: Sellers
authors.forename: Francesco
authors.initials: F
authors.lastname: Hofmann
mesh_terms: D019943:Amino Acid Substitution; D000631:Aminopyridines / Q000378:metabolism / Q000494:pharmacology; D000818:Animals; D000970:Antineoplastic Agents / Q000378:metabolism / Q000494:pharmacology*; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000378:metabolism / Q000494:pharmacology*; D002461:Cell Line, Transformed; D045744:Cell Line, Tumor; D018360:Crystallography, X-Ray; D004252:DNA Mutational Analysis; D004273:DNA, Neoplasm / Q000235:genetics; D019008:Drug Resistance, Neoplasm / Q000235:genetics*; D004789:Enzyme Activation / Q000235:genetics; D006801:Humans; D051379:Mice; D008958:Models, Molecular; D016296:Mutagenesis; D020125:Mutation, Missense; D009369:Neoplasms / Q000188:drug therapy / Q000235:genetics; D017354:Point Mutation; D011485:Protein Binding; D011487:Protein Conformation; D047428:Protein Kinase Inhibitors / Q000378:metabolism / Q000494:pharmacology*; D017434:Protein Structure, Tertiary; D019859:Proto-Oncogene Proteins c-met / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics; D011720:Pyrazoles / Q000378:metabolism / Q000494:pharmacology; D011804:Quinolines / Q000378:metabolism / Q000494:pharmacology*; D017978:Receptors, Growth Factor / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics; D014443:Tyrosine / Q000378:metabolism
chemical_list: C530596:1-(2-hydroxy-2-methylpropyl)-N-(5-(7-methoxyquinolin-4-yloxy)pyridin-2-yl)-5-methyl-3-oxo-2-phenyl-2,3-dihydro-1H-pyrazole-4-carboxamide; C561499:6-(6-(1-methyl-1H-pyrazol-4-yl)imidazo(1,2-b)pyridazin-3-ylmethyl)quinoline; D000631:Aminopyridines; D000970:Antineoplastic Agents; D019086:Bridged Bicyclo Compounds, Heterocyclic; D004273:DNA, Neoplasm; D047428:Protein Kinase Inhibitors; D011720:Pyrazoles; D011804:Quinolines; D017978:Receptors, Growth Factor; D014443:Tyrosine; C491743:MET protein, human; D019859:Proto-Oncogene Proteins c-met