title: Covalent and allosteric inhibitors of the ATPase VCP/p97 induce cancer cell death.
abstract: VCP (also known as p97 or Cdc48p in yeast) is an AAA(+) ATPase regulating endoplasmic reticulum-associated degradation. After high-throughput screening, we developed compounds that inhibit VCP via different mechanisms, including covalent modification of an active site cysteine and a new allosteric mechanism. Using photoaffinity labeling, structural analysis and mutagenesis, we mapped the binding site of allosteric inhibitors to a region spanning the D1 and D2 domains of adjacent protomers encompassing elements important for nucleotide-state sensing and ATP hydrolysis. These compounds induced an increased affinity for nucleotides. Interference with nucleotide turnover in individual subunits and distortion of interprotomer communication cooperated to impair VCP enzymatic activity. Chemical expansion of this allosteric class identified NMS-873, the most potent and specific VCP inhibitor described to date, which activated the unfolded protein response, interfered with autophagy and induced cancer cell death. The consistent pattern of cancer cell killing by covalent and allosteric inhibitors provided critical validation of VCP as a cancer target.
authors.affiliation: Business Unit Oncology, Nerviano Medical Sciences, Nerviano, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Magnaghi
authors.forename: Roberto
authors.initials: R
authors.lastname: D'Alessio
authors.forename: Barbara
authors.initials: B
authors.lastname: Valsasina
authors.forename: Nilla
authors.initials: N
authors.lastname: Avanzi
authors.forename: Simona
authors.initials: S
authors.lastname: Rizzi
authors.forename: Daniela
authors.initials: D
authors.lastname: Asa
authors.forename: Fabio
authors.initials: F
authors.lastname: Gasparri
authors.forename: Liviana
authors.initials: L
authors.lastname: Cozzi
authors.forename: Ulisse
authors.initials: U
authors.lastname: Cucchi
authors.forename: Christian
authors.initials: C
authors.lastname: Orrenius
authors.forename: Paolo
authors.initials: P
authors.lastname: Polucci
authors.forename: Dario
authors.initials: D
authors.lastname: Ballinari
authors.forename: Claudia
authors.initials: C
authors.lastname: Perrera
authors.forename: Antonella
authors.initials: A
authors.lastname: Leone
authors.forename: Giovanni
authors.initials: G
authors.lastname: Cervi
authors.forename: Elena
authors.initials: E
authors.lastname: Casale
authors.forename: Yang
authors.initials: Y
authors.lastname: Xiao
authors.forename: Chihunt
authors.initials: C
authors.lastname: Wong
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Anderson
authors.forename: Arturo
authors.initials: A
authors.lastname: Galvani
authors.forename: Daniele
authors.initials: D
authors.lastname: Donati
authors.forename: Tom
authors.initials: T
authors.lastname: O'Brien
authors.forename: Peter K
authors.initials: PK
authors.lastname: Jackson
authors.forename: Antonella
authors.initials: A
authors.lastname: Isacchi
mesh_terms: D000083:Acetanilides / Q000737:chemistry / Q000494:pharmacology*; D000251:Adenosine Triphosphatases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D000494:Allosteric Regulation / Q000187:drug effects; D000970:Antineoplastic Agents / Q000737:chemistry / Q000494:pharmacology*; D052160:Benzothiazoles / Q000737:chemistry / Q000494:pharmacology*; D018797:Cell Cycle Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects; D045744:Cell Line, Tumor; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D006801:Humans; D008958:Models, Molecular; D015394:Molecular Structure; D009369:Neoplasms / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology*; D013329:Structure-Activity Relationship; D000074405:Valosin Containing Protein
chemical_list: D000083:Acetanilides; D000970:Antineoplastic Agents; D052160:Benzothiazoles; D018797:Cell Cycle Proteins; D004791:Enzyme Inhibitors; C000623013:NMS-873; D000251:Adenosine Triphosphatases; C000614178:VCP protein, human; D000074405:Valosin Containing Protein
references.citation: Proc Natl Acad Sci U S A. 2012 Jun 19;109(25):9792-7
references.pmid: 22675116
references.citation: Oncogene. 2010 Jan 21;29(3):451-62
references.pmid: 19881538
references.citation: Expert Rev Proteomics. 2006 Aug;3(4):399-408
references.pmid: 16901199
references.citation: Hum Mol Genet. 2010 Apr 15;19(R1):R38-45
references.pmid: 20410287
references.citation: J Mol Biol. 2005 Mar 25;347(2):437-52
references.pmid: 15740751
references.citation: Nat Cell Biol. 2011 Oct 23;13(11):1376-82
references.pmid: 22020440
references.citation: Mol Cancer Ther. 2009 Apr;8(4):921-9
references.pmid: 19372565
references.citation: Nat Cell Biol. 2011 Mar;13(3):184-90
references.pmid: 21364565
references.citation: Bioorg Med Chem Lett. 2010 Mar 1;20(5):1677-9
references.pmid: 20137940
references.citation: Autophagy. 2011 Sep;7(9):1091-2
references.pmid: 21606684
references.citation: J Cell Biol. 2009 Dec 14;187(6):875-88
references.pmid: 20008565
references.citation: J Biol Chem. 2008 Mar 21;283(12):7445-54
references.pmid: 18199748
references.citation: Proc Natl Acad Sci U S A. 2012 Mar 6;109(10):3737-41
references.pmid: 22355145
references.citation: Mol Biol Cell. 2011 Feb 1;22(3):291-300
references.pmid: 21118995
references.citation: Nat Cell Biol. 2001 Aug;3(8):740-4
references.pmid: 11483959
references.citation: Genes Cells. 2010 Aug;15(8):911-22
references.pmid: 20604808
references.citation: Trends Biochem Sci. 2007 Jan;32(1):6-11
references.pmid: 17142044
references.citation: Mol Cell. 2011 Jun 24;42(6):731-43
references.pmid: 21700220
references.citation: Bioorg Med Chem Lett. 2011 May 15;21(10):2969-74
references.pmid: 21470862
references.citation: Nat Struct Biol. 2002 Dec;9(12):950-7
references.pmid: 12434150
references.citation: Nature. 2007 Jun 14;447(7146):859-63
references.pmid: 17568747
references.citation: Autophagy. 2010 Feb;6(2):217-27
references.pmid: 20104022
references.citation: J Biol Chem. 2008 May 16;283(20):13745-52
references.pmid: 18332143
references.citation: Clin Cancer Res. 2004 May 1;10(9):3007-12
references.pmid: 15131036
references.citation: Proc Natl Acad Sci U S A. 2008 Sep 2;105(35):12879-84
references.pmid: 18728180
references.citation: Semin Cell Dev Biol. 2007 Dec;18(6):780-91
references.pmid: 17942349
references.citation: Proc Natl Acad Sci U S A. 2011 Mar 22;108(12):4834-9
references.pmid: 21383145
references.citation: J Struct Biol. 2003 Dec;144(3):337-48
references.pmid: 14643202
references.citation: Biochem Biophys Res Commun. 2007 Mar 9;354(2):459-65
references.pmid: 17239345
references.citation: EMBO J. 2006 Jul 26;25(14):3357-66
references.pmid: 16810319
references.citation: Genes Dev. 2004 Dec 15;18(24):3066-77
references.pmid: 15601821
references.citation: Mol Cell Biol. 2009 Aug;29(16):4484-94
references.pmid: 19506019
references.citation: Genes Cells. 2001 Jul;6(7):575-97
references.pmid: 11473577
references.citation: Biochim Biophys Acta. 1995 Nov 15;1253(1):25-32
references.pmid: 7492595
references.citation: J Med Chem. 2013 Jan 24;56(2):437-50
references.pmid: 23245311
references.citation: PLoS Biol. 2006 Nov;4(11):e374
references.pmid: 17090218
references.citation: Oncogene. 2010 Aug 5;29(31):4424-35
references.pmid: 20514020
references.citation: J Biol Chem. 2005 Dec 16;280(50):41332-41
references.pmid: 16234241
references.citation: Structure. 2008 May;16(5):715-26
references.pmid: 18462676
references.citation: Jpn J Cancer Res. 2002 Mar;93(3):296-304
references.pmid: 11927012
references.citation: FEBS Lett. 2006 Jan 23;580(2):474-8
references.pmid: 16386250
references.citation: J Cell Sci. 2004 Jan 15;117(Pt 2):281-92
references.pmid: 14657277
references.citation: ChemMedChem. 2013 Feb;8(2):297-312
references.pmid: 23316025
references.citation: Cell. 2008 Sep 5;134(5):804-16
references.pmid: 18775313
references.citation: PLoS Biol. 2006 Nov;4(12):e423
references.pmid: 17132049
references.citation: EMBO J. 2010 Mar 3;29(5):969-80
references.pmid: 20075865
references.citation: Mol Biol Cell. 2006 Nov;17(11):4606-18
references.pmid: 16914519
references.citation: EMBO J. 2006 May 3;25(9):1967-76
references.pmid: 16601695