title: Multidomain targeting of Bcr-Abl by disruption of oligomerization and tyrosine kinase inhibition: toward eradication of CML.
abstract: The oncoprotein Bcr-Abl, the causative agent of chronic myeloid leukemia (CML), requires homo-oligomerization via a coiled-coil domain to function [Bartram, C. R.; et al. Nature 1983, 306 (5940), 277-280; and Zhao, X.; et al. Nat. Struct. Biol. 2002, 9(2), 117-120]. While tyrosine kinase inhibitors (TKIs) have shown great efficacy as treatment options for CML, their use may cause an acquisition of mutations in the tyrosine kinase domain, which prevent TKI binding and lead to a loss in activity [Woessner, D. W.; et al. Cancer J. 2011, 17(6), 477-486]. Previously, we have shown that a rationally modified coiled-coil domain (CC(mut3)) can disrupt this oligomerization, inhibit proliferation, and induce apoptosis in CML cells [Dixon, A. S.; et al. Mol. Pharmaceutics 2012, 9(1), 187-195]. Here, we show that using the most recently approved TKI, ponatinib (Iclusig), in combination with CC(mut3) allows a dose reduction of ponatinib and increased therapeutic efficacy in vitro measured by reduction in kinase activity, induction of apoptosis via caspase-3/7 and 7-AAD/Annexin V assays, and reduced transformative ability measured by a colony forming assay. The combination was effective not only in cells containing wild-type Bcr-Abl (K562, Ba/F3-p210) but also cells with Bcr-Abl containing the T315I mutation (Ba/F3-p210-T315I). In addition, we report for the first time the ability of CC(mut3) alone to inhibit the T315I mutant form of Bcr-Abl. This novel combination may prove to be more potent than single agent therapies and should be further explored for clinical use.
authors.affiliation: Department of Pharmaceutics and Pharmaceutical Chemistry, ‡Department of Pharmacology and Toxicology, College of Pharmacy, University of Utah , Salt Lake City, Utah 84112, United States.
authors.forename: Geoffrey D
authors.initials: GD
authors.lastname: Miller
authors.forename: David W
authors.initials: DW
authors.lastname: Woessner
authors.forename: Monika J
authors.initials: MJ
authors.lastname: Sirch
authors.forename: Carol S
authors.initials: CS
authors.lastname: Lim
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D053148:Caspase 3 / Q000378:metabolism; D053179:Caspase 7 / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects; D016044:Fusion Proteins, bcr-abl / Q000037:antagonists & inhibitors*; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology*; D020014:K562 Cells; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000378:metabolism*; D009154:Mutation; D009336:Necrosis / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D011724:Pyridazines / Q000494:pharmacology*
chemical_list: D007093:Imidazoles; D047428:Protein Kinase Inhibitors; D011724:Pyridazines; C545373:ponatinib; D016044:Fusion Proteins, bcr-abl; D053148:Caspase 3; D053179:Caspase 7
references.citation: J Biol Chem. 1996 Sep 13;271(37):22823-30
references.pmid: 8798460
references.citation: Oncogene. 1993 Jan;8(1):101-9
references.pmid: 8423987
references.citation: Mol Cancer. 2010 Apr 12;9:75
references.pmid: 20385023
references.citation: Leukemia. 2009 Dec;23(12):2242-7
references.pmid: 19798092
references.citation: Nature. 1983 Nov 17-23;306(5940):277-80
references.pmid: 6580527
references.citation: Oncogene. 2002 Dec 9;21(56):8629-42
references.pmid: 12476309
references.citation: Cancer Control. 2009 Apr;16(2):100-7
references.pmid: 19337196
references.citation: Nat Rev Cancer. 2012 Jul 24;12(8):513-26
references.pmid: 22825216
references.citation: Cytokine Growth Factor Rev. 1999 Jun;10(2):131-57
references.pmid: 10743504
references.citation: Blood. 2013 Jan 17;121(3):489-98
references.pmid: 23223358
references.citation: Mol Cell Biol. 1993 Dec;13(12):7587-95
references.pmid: 8246975
references.citation: PLoS One. 2011 Feb 11;6(2):e17020
references.pmid: 21347248
references.citation: Cancer Res. 2002 Dec 15;62(24):7149-53
references.pmid: 12499247
references.citation: J Biol Chem. 2011 Aug 5;286(31):27751-60
references.pmid: 21659527
references.citation: J Immunol. 1982 Jun;128(6):2393-8
references.pmid: 6122701
references.citation: Leukemia. 2012 Feb;26(2):214-24
references.pmid: 21844872
references.citation: Eur J Haematol. 2004 Jun;72(6):420-9
references.pmid: 15128421
references.citation: Nat Rev Clin Oncol. 2013 Feb;10(2):65
references.pmid: 23229179
references.citation: Mol Pharm. 2013 Jan 7;10(1):270-7
references.pmid: 23211037
references.citation: J Biol Chem. 2001 Oct 19;276(42):39462-8
references.pmid: 11502748
references.citation: Cancer Cell. 2009 Nov 6;16(5):401-12
references.pmid: 19878872
references.citation: Science. 1986 Jul 11;233(4760):212-4
references.pmid: 3460176
references.citation: Pharm Res. 2012 Apr;29(4):1098-109
references.pmid: 22183511
references.citation: Nature. 1973 Jun 1;243(5405):290-3
references.pmid: 4126434
references.citation: N Engl J Med. 2012 Nov 29;367(22):2075-88
references.pmid: 23190221
references.citation: Cancer J. 2011 Nov-Dec;17(6):477-86
references.pmid: 22157291
references.citation: Mol Cell. 2005 Aug 19;19(4):461-73
references.pmid: 16109371
references.citation: Haematologica. 2012 Sep;97(9):1444
references.pmid: 22532521
references.citation: Cell. 1991 Nov 1;67(3):459-68
references.pmid: 1657398
references.citation: J Med Chem. 2010 Jun 24;53(12):4701-19
references.pmid: 20513156
references.citation: Leukemia. 2010 Jan;24(1):226-9
references.pmid: 19759561
references.citation: Mol Cancer Ther. 2011 Jun;10(6):1028-35
references.pmid: 21482694
references.citation: J Cell Biol. 1994 Feb;124(3):325-40
references.pmid: 8294516
references.citation: Mol Pharm. 2012 Jan 1;9(1):187-95
references.pmid: 22136227
references.citation: Nat Rev Mol Cell Biol. 2004 Jan;5(1):33-44
references.pmid: 14708008
references.citation: Leukemia. 2008 Jun;22(6):1184-90
references.pmid: 18354488
references.citation: Blood. 2007 Oct 1;110(7):2242-9
references.pmid: 17496200
references.citation: Nat Struct Biol. 2002 Feb;9(2):117-20
references.pmid: 11780146