title: Crystal structure of the FLT3 kinase domain bound to the inhibitor Quizartinib (AC220).
abstract: More than 30% of acute myeloid leukemia (AML) patients possess activating mutations in the receptor tyrosine kinase FMS-like tyrosine kinase 3 or FLT3. A small-molecule inhibitor of FLT3 (known as quizartinib or AC220) that is currently in clinical trials appears promising for the treatment of AML. Here, we report the co-crystal structure of the kinase domain of FLT3 in complex with quizartinib. FLT3 with quizartinib bound adopts an "Abl-like" inactive conformation with the activation loop stabilized in the "DFG-out" orientation and folded back onto the kinase domain. This conformation is similar to that observed for the uncomplexed intracellular domain of FLT3 as well as for related receptor tyrosine kinases, except for a localized induced fit in the activation loop. The co-crystal structure reveals the interactions between quizartinib and the active site of FLT3 that are key for achieving its high potency against both wild-type FLT3 as well as a FLT3 variant observed in many AML patients. This co-complex further provides a structural rationale for quizartinib-resistance mutations.
authors.affiliation: Department of Molecular and Cell Biology, University of California, Berkeley, California, United States of America; California Institute for Quantitative Biosciences, University of California, Berkeley, California, United States of America.
authors.forename: Julie A
authors.initials: JA
authors.lastname: Zorn
authors.affiliation: Department of Molecular and Cell Biology, University of California, Berkeley, California, United States of America; California Institute for Quantitative Biosciences, University of California, Berkeley, California, United States of America.
authors.forename: Qi
authors.initials: Q
authors.lastname: Wang
authors.affiliation: Department of Molecular and Cell Biology, University of California, Berkeley, California, United States of America.
authors.forename: Eric
authors.initials: E
authors.lastname: Fujimura
authors.affiliation: Department of Molecular and Cell Biology, University of California, Berkeley, California, United States of America; California Institute for Quantitative Biosciences, University of California, Berkeley, California, United States of America; Howard Hughes Medical Institute, University of California, Berkeley, California, United States of America.
authors.forename: Tiago
authors.initials: T
authors.lastname: Barros
authors.affiliation: Department of Molecular and Cell Biology, University of California, Berkeley, California, United States of America; California Institute for Quantitative Biosciences, University of California, Berkeley, California, United States of America; Howard Hughes Medical Institute, University of California, Berkeley, California, United States of America; Department of Chemistry, University of California, Berkeley, California, United States of America; Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, California, United States of America.
authors.forename: John
authors.initials: J
authors.lastname: Kuriyan
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000737:chemistry*; D016367:Baculoviridae / Q000235:genetics; D052160:Benzothiazoles / Q000737:chemistry*; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D006801:Humans; D062105:Molecular Docking Simulation; D056004:Molecular Dynamics Simulation; D009154:Mutation; D009363:Neoplasm Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics; D010671:Phenylurea Compounds / Q000737:chemistry*; D011485:Protein Binding; D017510:Protein Folding; D047428:Protein Kinase Inhibitors / Q000737:chemistry*; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000235:genetics; D061987:Sf9 Cells; D018411:Spodoptera / Q000235:genetics; D051941:fms-Like Tyrosine Kinase 3 / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics
chemical_list: D000970:Antineoplastic Agents; D052160:Benzothiazoles; D009363:Neoplasm Proteins; D010671:Phenylurea Compounds; D047428:Protein Kinase Inhibitors; D011993:Recombinant Fusion Proteins; C544967:quizartinib; C497970:FLT3 protein, human; D051941:fms-Like Tyrosine Kinase 3
references.citation: Proteins. 2010 Jun;78(8):1950-8
references.pmid: 20408171
references.citation: Cell. 2010 Jun 25;141(7):1117-34
references.pmid: 20602996
references.citation: Annu Rev Biochem. 2006;75:93-109
references.pmid: 16756486
references.citation: Proc Natl Acad Sci U S A. 2014 Apr 8;111(14):5319-24
references.pmid: 24623852
references.citation: Methods Enzymol. 2014;548:23-67
references.pmid: 25399641
references.citation: Bioorg Chem. 2011 Dec;39(5-6):192-210
references.pmid: 21872901
references.citation: J Med Chem. 2009 Dec 10;52(23):7808-16
references.pmid: 19754199
references.citation: Bioinformatics. 2013 Apr 1;29(7):845-54
references.pmid: 23407358
references.citation: Nature. 2001 May 17;411(6835):355-65
references.pmid: 11357143
references.citation: Blood. 2011 Jul 7;118(1):60-8
references.pmid: 21389326
references.citation: Leukemia. 2013 Jun;27(6):1416-8
references.pmid: 23392356
references.citation: Int J Hematol. 2005 Aug;82(2):100-7
references.pmid: 16146839
references.citation: Science. 2000 Sep 15;289(5486):1938-42
references.pmid: 10988075
references.citation: Cancer Res. 2006 Jan 15;66(2):1007-14
references.pmid: 16424036
references.citation: ACS Chem Biol. 2013 Jan 18;8(1):96-104
references.pmid: 23276252
references.citation: Mol Cancer Ther. 2013 Apr;12(4):438-47
references.pmid: 23412931
references.citation: Nat Rev Mol Cell Biol. 2004 Jun;5(6):464-71
references.pmid: 15173825
references.citation: Oncogene. 2000 Aug 17;19(35):3961-70
references.pmid: 10962552
references.citation: Blood. 2002 Sep 1;100(5):1532-42
references.pmid: 12176867
references.citation: Cell. 2002 May 3;109(3):275-82
references.pmid: 12015977
references.citation: J Biol Chem. 2004 Jul 23;279(30):31655-63
references.pmid: 15123710
references.citation: Biochemistry. 2009 Jul 28;48(29):7019-31
references.pmid: 19526984
references.citation: Methods Enzymol. 1997;276:307-26
references.pmid: 27754618
references.citation: Curr Hematol Malig Rep. 2014 Jun;9(2):174-85
references.pmid: 24682858
references.citation: Acta Crystallogr D Biol Crystallogr. 2010 Feb;66(Pt 2):213-21
references.pmid: 20124702
references.citation: Mol Cell. 1999 May;3(5):639-48
references.pmid: 10360180
references.citation: Nature. 2012 Apr 15;485(7397):260-3
references.pmid: 22504184
references.citation: Bioorg Med Chem Lett. 2009 Sep 1;19(17):5182-5
references.pmid: 19646870
references.citation: Cell. 2001 Sep 21;106(6):745-57
references.pmid: 11572780
references.citation: Mol Cell. 2004 Jan 30;13(2):169-78
references.pmid: 14759363
references.citation: ACS Chem Biol. 2014 Jun 20;9(6):1230-41
references.pmid: 24730530
references.citation: Nat Chem Biol. 2006 Jul;2(7):358-64
references.pmid: 16783341
references.citation: Cold Spring Harb Perspect Biol. 2014 Mar 01;6(3):null
references.pmid: 24591517
references.citation: J Biol Chem. 2003 Aug 22;278(34):31461-4
references.pmid: 12824176
references.citation: Blood. 2009 Oct 1;114(14 ):2984-92
references.pmid: 19654408
references.citation: Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2126-32
references.pmid: 15572765
references.citation: Proc Natl Acad Sci U S A. 2012 Nov 6;109(45):18281-9
references.pmid: 22988103