title: Development of covalent inhibitors that can overcome resistance to first-generation FGFR kinase inhibitors.
abstract: The human FGF receptors (FGFRs) play critical roles in various human cancers, and several FGFR inhibitors are currently under clinical investigation. Resistance usually results from selection for mutant kinases that are impervious to the action of the drug or from up-regulation of compensatory signaling pathways. Preclinical studies have demonstrated that resistance to FGFR inhibitors can be acquired through mutations in the FGFR gatekeeper residue, as clinically observed for FGFR4 in embryonal rhabdomyosarcoma and neuroendocrine breast carcinomas. Here we report on the use of a structure-based drug design to develop two selective, next-generation covalent FGFR inhibitors, the FGFR irreversible inhibitors 2 (FIIN-2) and 3 (FIIN-3). To our knowledge, FIIN-2 and FIIN-3 are the first inhibitors that can potently inhibit the proliferation of cells dependent upon the gatekeeper mutants of FGFR1 or FGFR2, which confer resistance to first-generation clinical FGFR inhibitors such as NVP-BGJ398 and AZD4547. Because of the conformational flexibility of the reactive acrylamide substituent, FIIN-3 has the unprecedented ability to inhibit both the EGF receptor (EGFR) and FGFR covalently by targeting two distinct cysteine residues. We report the cocrystal structure of FGFR4 with FIIN-2, which unexpectedly exhibits a "DFG-out" covalent binding mode. The structural basis for dual FGFR and EGFR targeting by FIIN3 also is illustrated by crystal structures of FIIN-3 bound with FGFR4 V550L and EGFR L858R. These results have important implications for the design of covalent FGFR inhibitors that can overcome clinical resistance and provide the first example, to our knowledge, of a kinase inhibitor that covalently targets cysteines located in different positions within the ATP-binding pocket.
authors.affiliation: Departments of Biological Chemistry and Molecular Pharmacology, Departments of Cancer Biology and.
authors.forename: Li
authors.initials: L
authors.lastname: Tan
authors.affiliation: Medical Oncology.
authors.forename: Jun
authors.initials: J
authors.lastname: Wang
authors.affiliation: Medical Oncology, The Lowe Center for Thoracic Oncology.
authors.forename: Junko
authors.initials: J
authors.lastname: Tanizaki
authors.affiliation: Department of Biochemistry and Molecular Pharmacology, New York University School of Medicine, New York, NY 10016; School of Pharmacy, Wenzhou Medical University, Wenzhou, 325035, China;
authors.forename: Zhifeng
authors.initials: Z
authors.lastname: Huang
authors.affiliation: Medical Oncology, Genetics, and.
authors.forename: Amir R
authors.initials: AR
authors.lastname: Aref
authors.affiliation: Medical Oncology, Department of Clinical Medicine, Aarhus University, Aarhus, 8200 Denmark;
authors.forename: Maria
authors.initials: M
authors.lastname: Rusan
authors.affiliation: Peking University Health Science Center, Beijing, 100191, China;
authors.forename: Su-Jie
authors.initials: SJ
authors.lastname: Zhu
authors.affiliation: Medicine, Harvard Medical School, Boston, MA 02115; Cardiovascular Research Center, Massachusetts General Hospital, Charlestown, MA 02129;
authors.forename: Yiyun
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Medical Oncology, The Lowe Center for Thoracic Oncology.
authors.forename: Dalia
authors.initials: D
authors.lastname: Ercan
authors.affiliation: Medical Oncology, Cancer Program, Broad Institute of Harvard and MIT, Cambridge, MA 02141;
authors.forename: Rachel G
authors.initials: RG
authors.lastname: Liao
authors.affiliation: Medical Oncology, The Lowe Center for Thoracic Oncology.
authors.forename: Marzia
authors.initials: M
authors.lastname: Capelletti
authors.affiliation: Departments of Biological Chemistry and Molecular Pharmacology, Departments of Cancer Biology and.
authors.forename: Wenjun
authors.initials: W
authors.lastname: Zhou
authors.affiliation: Departments of Biological Chemistry and Molecular Pharmacology, Departments of Cancer Biology and Chemical Kinomics Research Center, Korea Institute of Science and Technology, Seoul, 136-791 Republic of Korea;
authors.forename: Wooyoung
authors.initials: W
authors.lastname: Hur
authors.affiliation: New Drug Development Center, Daegu-Gyeongbuk Medical Innovation Foundation, Daegu, 706-010 Republic of Korea; and.
authors.forename: NamDoo
authors.initials: N
authors.lastname: Kim
authors.affiliation: Chemical Kinomics Research Center, Korea Institute of Science and Technology, Seoul, 136-791 Republic of Korea; Korea University-Korean Institute of Science and Technology Graduate School of Converging Science and Technology, Seoul, 136-713 Republic of Korea.
authors.forename: Taebo
authors.initials: T
authors.lastname: Sim
authors.affiliation: Departments of Cancer Biology and Genetics, and Center for Systems Cancer Biology, and.
authors.forename: Suzanne
authors.initials: S
authors.lastname: Gaudet
authors.affiliation: Medical Oncology.
authors.forename: David A
authors.initials: DA
authors.lastname: Barbie
authors.affiliation: Medicine, Harvard Medical School, Boston, MA 02115; Cardiovascular Research Center, Massachusetts General Hospital, Charlestown, MA 02129;
authors.forename: Jing-Ruey Joanna
authors.initials: JR
authors.lastname: Yeh
authors.affiliation: Peking University Health Science Center, Beijing, 100191, China;
authors.forename: Cai-Hong
authors.initials: CH
authors.lastname: Yun
authors.affiliation: Medical Oncology, Cancer Program, Broad Institute of Harvard and MIT, Cambridge, MA 02141; Peter_Hammerman@dfci.harvard.edu Moosa.Mohammadi@nyumc.org pasi_janne@dfci.harvard.edu nathanael_gray@dfci.harvard.edu.
authors.forename: Peter S
authors.initials: PS
authors.lastname: Hammerman
authors.affiliation: Department of Biochemistry and Molecular Pharmacology, New York University School of Medicine, New York, NY 10016; Peter_Hammerman@dfci.harvard.edu Moosa.Mohammadi@nyumc.org pasi_janne@dfci.harvard.edu nathanael_gray@dfci.harvard.edu.
authors.forename: Moosa
authors.initials: M
authors.lastname: Mohammadi
authors.affiliation: Medical Oncology, The Lowe Center for Thoracic Oncology, Belfer Institute for Applied Cancer Science, Dana Farber Cancer Institute, Boston, MA 02215; Peter_Hammerman@dfci.harvard.edu Moosa.Mohammadi@nyumc.org pasi_janne@dfci.harvard.edu nathanael_gray@dfci.harvard.edu.
authors.forename: Pasi A
authors.initials: PA
authors.lastname: Jänne
authors.affiliation: Departments of Biological Chemistry and Molecular Pharmacology, Departments of Cancer Biology and Peter_Hammerman@dfci.harvard.edu Moosa.Mohammadi@nyumc.org pasi_janne@dfci.harvard.edu nathanael_gray@dfci.harvard.edu.
authors.forename: Nathanael S
authors.initials: NS
authors.lastname: Gray
mesh_terms: D019943:Amino Acid Substitution; D000970:Antineoplastic Agents / Q000737:chemistry / Q000494:pharmacology; D001665:Binding Sites; D045744:Cell Line, Tumor; D018360:Crystallography, X-Ray; D019008:Drug Resistance, Neoplasm / Q000187:drug effects* / Q000235:genetics; D066246:ErbB Receptors / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006801:Humans; D020125:Mutation, Missense; D009369:Neoplasms / Q000188:drug therapy* / Q000201:enzymology / Q000235:genetics / Q000473:pathology; D047428:Protein Kinase Inhibitors / Q000737:chemistry / Q000494:pharmacology; D051496:Receptor, Fibroblast Growth Factor, Type 1 / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D051497:Receptor, Fibroblast Growth Factor, Type 2 / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D051499:Receptor, Fibroblast Growth Factor, Type 4 / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013329:Structure-Activity Relationship
chemical_list: D000970:Antineoplastic Agents; D047428:Protein Kinase Inhibitors; C512478:EGFR protein, human; D066246:ErbB Receptors; C496348:FGFR1 protein, human; C496353:FGFR2 protein, human; C496371:FGFR4 protein, human; D051496:Receptor, Fibroblast Growth Factor, Type 1; D051497:Receptor, Fibroblast Growth Factor, Type 2; D051499:Receptor, Fibroblast Growth Factor, Type 4
keywords: cancer drug resistance; drug discovery; kinase inhibitor; structure-based drug design