Targeting Drug Resistance in EGFR with Covalent Inhibitors: A Structure-Based Design Approach.
Receptor tyrosine kinases represent one of the prime targets in cancer therapy, as the dysregulation of these elementary transducers of extracellular signals, like the epidermal growth factor receptor (EGFR), contributes to the onset of cancer, such as non-small cell lung cancer (NSCLC). Strong efforts were directed to the development of irreversible inhibitors and led to compound CO-1686, which takes advantage of increased residence time at EGFR by alkylating Cys797 and thereby preventing toxic effects. Here, we present a structure-based approach, rationalized by subsequent computational analysis of conformational ligand ensembles in solution, to design novel and irreversible EGFR inhibitors based on a screening hit that was identified in a phenotype screen of 80 NSCLC cell lines against approximately 1500 compounds. Using protein X-ray crystallography, we deciphered the binding mode in engineered cSrc (T338M/S345C), a validated model system for EGFR-T790M, which constituted the basis for further rational design approaches. Chemical synthesis led to further compound collections that revealed increased biochemical potency and, in part, selectivity toward mutated (L858R and L858R/T790M) vs nonmutated EGFR. Further cell-based and kinetic studies were performed to substantiate our initial findings. Utilizing proteolytic digestion and nano-LC-MS/MS analysis, we confirmed the alkylation of Cys797.
Department of Chemistry and Chemical Biology, TU Dortmund University , Otto-Hahn-Straße 6, D-44227 Dortmund, Germany.
Julian
J
Engel
Department of Chemistry and Chemical Biology, TU Dortmund University , Otto-Hahn-Straße 6, D-44227 Dortmund, Germany.
André
A
Richters
Chemical Genomics Centre of the Max-Planck Society , Otto-Hahn-Straße 15, D-44227 Dortmund, Germany.
Matthäus
M
Getlik
Department of Chemistry and Chemical Biology, TU Dortmund University , Otto-Hahn-Straße 6, D-44227 Dortmund, Germany.
Stefano
S
Tomassi
Department of Chemistry and Chemical Biology, TU Dortmund University , Otto-Hahn-Straße 6, D-44227 Dortmund, Germany.
Marina
M
Keul
Chemical Genomics Centre of the Max-Planck Society , Otto-Hahn-Straße 15, D-44227 Dortmund, Germany.
Martin
M
Termathe
Department of Chemistry and Chemical Biology, TU Dortmund University , Otto-Hahn-Straße 6, D-44227 Dortmund, Germany.
Jonas
J
Lategahn
Department of Chemistry and Chemical Biology, TU Dortmund University , Otto-Hahn-Straße 6, D-44227 Dortmund, Germany.
Christian
C
Becker
Department of Chemistry and Chemical Biology, TU Dortmund University , Otto-Hahn-Straße 6, D-44227 Dortmund, Germany.
Svenja
S
Mayer-Wrangowski
Department of Chemistry and Chemical Biology, TU Dortmund University , Otto-Hahn-Straße 6, D-44227 Dortmund, Germany.
Christian
C
Grütter
Department of Chemistry and Chemical Biology, TU Dortmund University , Otto-Hahn-Straße 6, D-44227 Dortmund, Germany.
Niklas
N
Uhlenbrock
Department of Chemistry and Chemical Biology, TU Dortmund University , Otto-Hahn-Straße 6, D-44227 Dortmund, Germany.
Jasmin
J
Krüll
Department of Chemistry and Chemical Biology, TU Dortmund University , Otto-Hahn-Straße 6, D-44227 Dortmund, Germany.
Niklas
N
Schaumann
Department of Chemistry and Chemical Biology, TU Dortmund University , Otto-Hahn-Straße 6, D-44227 Dortmund, Germany.
Simone
S
Eppmann
Department of Chemistry and Chemical Biology, TU Dortmund University , Otto-Hahn-Straße 6, D-44227 Dortmund, Germany.
Patrick
P
Kibies
Department of Chemistry and Chemical Biology, TU Dortmund University , Otto-Hahn-Straße 6, D-44227 Dortmund, Germany.
Franziska
F
Hoffgaard
Department of Chemistry and Chemical Biology, TU Dortmund University , Otto-Hahn-Straße 6, D-44227 Dortmund, Germany.
Jochen
J
Heil
Lead Discovery Center GmbH , Otto-Hahn-Straße 15, D-44227 Dortmund, Germany.
Sascha
S
Menninger
Department of Translational Genomics, Medical Faculty, University of Cologne , Weyertal 115b, D-50931 Cologne, Germany.
Sandra
S
Ortiz-Cuaran
Department of Translational Genomics, Medical Faculty, University of Cologne , Weyertal 115b, D-50931 Cologne, Germany.
Johannes M
JM
Heuckmann
Leibniz-Institut für Analytische Wissenschaften - ISAS - e.V. , D-44139 Dortmund, Germany.
Verena
V
Tinnefeld
Leibniz-Institut für Analytische Wissenschaften - ISAS - e.V. , D-44139 Dortmund, Germany.
René P
RP
Zahedi
Department of Translational Genomics, Medical Faculty, University of Cologne , Weyertal 115b, D-50931 Cologne, Germany.|Molecular Pathology, University Hospital of Cologne , Kerpenerstraße 62, D-50937 Cologne, Germany.
Martin L
ML
Sos
Lead Discovery Center GmbH , Otto-Hahn-Straße 15, D-44227 Dortmund, Germany.
Carsten
C
Schultz-Fademrecht
Department of Translational Genomics, Medical Faculty, University of Cologne , Weyertal 115b, D-50931 Cologne, Germany.|Department of Pathology, University of Cologne , Joseph-Stelzmann Straße 9, D-50931 Cologne, Germany.
Roman K
RK
Thomas
Department of Chemistry and Chemical Biology, TU Dortmund University , Otto-Hahn-Straße 6, D-44227 Dortmund, Germany.
Stefan M
SM
Kast
Department of Chemistry and Chemical Biology, TU Dortmund University , Otto-Hahn-Straße 6, D-44227 Dortmund, Germany.|Chemical Genomics Centre of the Max-Planck Society , Otto-Hahn-Straße 15, D-44227 Dortmund, Germany.
Daniel
D
Rauh
D000970:Antineoplastic Agents / Q000737:chemistry* / Q000494:pharmacology; D002289:Carcinoma, Non-Small-Cell Lung; D045744:Cell Line, Tumor; D002463:Cell Membrane Permeability; D018360:Crystallography, X-Ray; D062126:Databases, Chemical; D015195:Drug Design; D019008:Drug Resistance, Neoplasm; D066246:ErbB Receptors / Q000037:antagonists & inhibitors* / Q000235:genetics; D006801:Humans; D007700:Kinetics; D008175:Lung Neoplasms; D008958:Models, Molecular; D008968:Molecular Conformation; D009154:Mutation; D011720:Pyrazoles / Q000737:chemistry / Q000494:pharmacology; D011743:Pyrimidines / Q000737:chemistry / Q000494:pharmacology; D011799:Quinazolines / Q000737:chemistry / Q000494:pharmacology; D054852:Small Molecule Libraries; D012995:Solubility; D013329:Structure-Activity Relationship; D019061:src-Family Kinases / Q000737:chemistry / Q000235:genetics
D000970:Antineoplastic Agents; D011720:Pyrazoles; D011743:Pyrimidines; D011799:Quinazolines; D054852:Small Molecule Libraries; C512478:EGFR protein, human; D066246:ErbB Receptors; D019061:src-Family Kinases