Targeting the Replication Checkpoint Using SCH 900776, a Potent and Functionally Selective CHK1 Inhibitor Identified via High Content Screening

Molecular Cancer Therapeutics - Tập 10 Số 4 - Trang 591-602 - 2011
Timothy J. Guzi1, Kamil Paruch1, Michael P. Dwyer1, Marc Labroli1, Fergus Shanahan1, Nicole R. Davis1, Lorena Taricani1, Derek Wiswell1, Wolfgang Seghezzi1, Ervin Penaflor1, Bhagyashree Bhagwat1, Wei Wang1, Danling Gu1, Yunsheng Hsieh1, Suining Lee1, Ming Liu1, David Parry1
1Authors' Affiliations: 1Merck Research Laboratory, Cambridge, Masachusetts; and 2Merck Research Laboratory, Kenilworth, New Jersey 3Merck Research Laboratory, Palo Alto, California

Tóm tắt

Abstract Checkpoint kinase 1 (CHK1) is an essential serine/threonine kinase that responds to DNA damage and stalled DNA replication. CHK1 is essential for maintenance of replication fork viability during exposure to DNA antimetabolites. In human tumor cell lines, ablation of CHK1 function during antimetabolite exposure led to accumulation of double-strand DNA breaks and cell death. Here, we extend these observations and confirm ablation of CHK2 does not contribute to these phenotypes and may diminish them. Furthermore, concomitant suppression of cyclin-dependent kinase (CDK) activity is sufficient to completely antagonize the desired CHK1 ablation phenotypes. These mechanism-based observations prompted the development of a high-content, cell-based screen for γ-H2AX induction, a surrogate marker for double-strand DNA breaks. This mechanism-based functional approach was used to optimize small molecule inhibitors of CHK1. Specifically, the assay was used to mechanistically define the optimal in-cell profile with compounds exhibiting varying degrees of CHK1, CHK2, and CDK selectivity. Using this approach, SCH 900776 was identified as a highly potent and functionally optimal CHK1 inhibitor with minimal intrinsic antagonistic properties. SCH 900776 exposure phenocopies short interfering RNA-mediated CHK1 ablation and interacts synergistically with DNA antimetabolite agents in vitro and in vivo to selectively induce dsDNA breaks and cell death in tumor cell backgrounds. Mol Cancer Ther; 10(4); 591–602. ©2011 AACR.

Từ khóa


Tài liệu tham khảo

Haskell, 2001, Cancer Treatment

Shi, 2001, S-Phase arrest by nucleoside analogues and abrogation of survival without cell cycle progression by 7-hydroxystaurosporine, Cancer Res, 61, 1065

Zhang, 2006, Turning the replication checkpoint on and off, Cell Cycle, 5, 125, 10.4161/cc.5.2.2308

Zachos, 2003, Chk1-deficient tumour cells are viable but exhibit multiple checkpoint and survival defects, EMBO J, 22, 713, 10.1093/emboj/cdg060

Cho, 2005, Chk1 is essential for tumor cell viability following activation of the replication checkpoint, Cell Cycle, 4, 131, 10.4161/cc.4.1.1299

Syljuasen, 2005, Inhibition of human Chk1 causes increased initiation of DNA replication, phosphorylation of ATR targets, and DNA breakage, Mol Cell Biol, 25, 3553, 10.1128/MCB.25.9.3553-3562.2005

Taricani, 2009, Replication stress activates DNA polymerase alpha-associated Chk1, Cell Cycle, 8, 482, 10.4161/cc.8.3.7661

Furnari, 1997, Cdc25 mitotic inducer targeted by chk1 DNA damage checkpoint kinase, Science, 277, 1495, 10.1126/science.277.5331.1495

O'Connell, 1997, Chk1 is a wee1 kinase in the G2 DNA damage checkpoint inhibiting cdc2 by Y15 phosphorylation, EMBO J, 16, 545, 10.1093/emboj/16.3.545

Sanchez, 1997, Conservation of the Chk1 checkpoint pathway in mammals: linkage of DNA damage to Cdk regulation through Cdc25, Science, 277, 1497, 10.1126/science.277.5331.1497

Liu, 2000, Chk1 is an essential kinase that is regulated by Atr and required for the G(2)/M DNA damage checkpoint, Genes Dev, 14, 1448, 10.1101/gad.14.12.1448

Ewald, 2007, H2AX phosphorylation marks gemcitabine-induced stalled replication forks and their collapse upon S-phase checkpoint abrogation, Mol Cancer Ther, 6, 1239, 10.1158/1535-7163.MCT-06-0633

Walton, 2010, The preclinical pharmacology and therapeutic activity of the novel CHK1 inhibitor SAR-020106, Mol Cancer Ther, 9, 89, 10.1158/1535-7163.MCT-09-0938

Dwyer, 2007, Versatile templates for the development of novel kinase inhibitors: discovery of novel CDK inhibitors, Bioorg Med Chem Lett, 17, 6216, 10.1016/j.bmcl.2007.09.018

Paruch, 2010, Discovery of dinaciclib (SCH 727965): a potent and selective inhibitor of cyclin-dependent kinases, ACS Med Chem Lett, 1, 204, 10.1021/ml100051d

Dwyer, 2010, Discovery of pyrazolo[1,5-a]pyrimidine-based CHK1 inhibitors: A template-based approach – part 1, Bioorg. Med. Chem. Lett., 10.1016/j.bmcl.2010.10.113

Labroli, 2010, Discovery of pyrazolo[1,5-a]pyrimidine-based CHK1 inhibitors: A template-based approach – part 2, Bioorg. Med. Chem. Lett., 10.1016/j.bmcl.2010.10.114

L'Italien, 2006, Unmasking the redundancy between Cdk1 and Cdk2 at G2 phase in human cancer cell lines, Cell Cycle, 5, 984, 10.4161/cc.5.9.2721

Paruch, 2007, Pyrazolo[1,5-a]pyrimidines as orally available inhibitors of cyclin-dependent kinase 2, Bioorg Med Chem Lett, 17, 6220, 10.1016/j.bmcl.2007.09.017

Parry, 2010, Dinaciclib (SCH 727965), a novel and potent cyclin-dependent kinase inhibitor, Mol Cancer Ther, 9, 2344, 10.1158/1535-7163.MCT-10-0324

Hsieh, 2009, The role of hyphenated chromatography-mass spectrometry techniques in exploratory drug metabolism and pharmacokinetics, Curr Pharm Des, 15, 2251, 10.2174/138161209788682505

Sampath, 2002, Inhibition of cyclin-dependent kinase 2 by the Chk1-Cdc25A pathway during the S-phase checkpoint activated by fludarabine: dysregulation by 7-hydroxystaurosporine, Mol Pharmacol, 62, 680, 10.1124/mol.62.3.680

Fischmann, 2008, Structure-guided discovery of cyclin-dependent kinase inhibitors, Biopolymers, 89, 372, 10.1002/bip.20868

Smits, 2006, Spreading the signal: dissociation of Chk1 from chromatin, Cell Cycle, 5, 1039, 10.4161/cc.5.10.2761

Clarke, 2005, DNA-dependent phosphorylation of Chk1 and Claspin in a human cell-free system, Biochem J, 388, 705, 10.1042/BJ20041966

Jardim, 2009, Reduced ATR or Chk1 expression leads to chromosome instability and chemosensitization of mismatch repair-deficient colorectal cancer cells, Mol Biol Cell, 20, 3801, 10.1091/mbc.e09-04-0303

Plunkett, 1996, Gemcitabine: preclinical pharmacology and mechanisms of action, Semin Oncol, 23, 3

Arumugam, 2009, Epithelial to mesenchymal transition contributes to drug resistance in pancreatic cancer, Cancer Res, 69, 5820, 10.1158/0008-5472.CAN-08-2819

Zabludoff, 2008, AZD7762, a novel checkpoint kinase inhibitor, drives checkpoint abrogation and potentiates DNA-targeted therapies, Mol Cancer Ther, 7, 2955, 10.1158/1535-7163.MCT-08-0492

Workman, 2003, How much gets there and what does it do?: The need for better pharmacokinetic and pharmacodynamic endpoints in contemporary drug discovery and development, Curr Pharm Des, 9, 891, 10.2174/1381612033455279

Feijoo, 2001, Activation of mammalian Chk1 during DNA replication arrest: a role for Chk1 in the intra-S phase checkpoint monitoring replication origin firing, J Cell Biol, 154, 913, 10.1083/jcb.200104099

McNeely, 2010, Chk1 inhibition after replicative stress activates a double strand break response mediated by ATM and DNA-dependent protein kinase, Cell Cycle, 995

Bartek, 2003, Chk1 and Chk2 kinases in checkpoint control and cancer, Cancer Cell, 3, 421, 10.1016/S1535-6108(03)00110-7

Wang, 1996, UCN-01: a potent abrogator of G2 checkpoint function in cancer cells with disrupted p53, J Natl Cancer Inst, 88, 956, 10.1093/jnci/88.14.956

Sugiyama, 2000, UCN-01 selectively enhances mitomycin C cytotoxicity in p53 defective cells which is mediated through S and/or G(2) checkpoint abrogation, Int J Cancer, 85, 703, 10.1002/(SICI)1097-0215(20000301)85:5<703::AID-IJC17>3.0.CO;2-7

Blasina, 2008, Breaching the DNA damage checkpoint via PF-00477736, a novel small-molecule inhibitor of checkpoint kinase 1, Mol Cancer Ther, 7, 2394, 10.1158/1535-7163.MCT-07-2391

Zhao, 2001, ATR-mediated checkpoint pathways regulate phosphorylation and activation of human Chk1, Mol Cell Biol, 21, 4129, 10.1128/MCB.21.13.4129-4139.2001

Lupardus, 2002, A requirement for replication in activation of the ATR-dependent DNA damage checkpoint, Genes Dev, 16, 2327, 10.1101/gad.1013502

Liu, 2006, Claspin operates downstream of TopBP1 to direct ATR signaling towards Chk1 activation, Mol Cell Biol, 26, 6056, 10.1128/MCB.00492-06

Paulsen, 2007, The ATR pathway: fine-tuning the fork, DNA Repair (Amst), 6, 953, 10.1016/j.dnarep.2007.02.015

Gandhi, 2002, Prolonged infusion of gemcitabine: clinical and pharmacodynamic studies during a phase I trial in relapsed acute myelogenous leukemia, J Clin Oncol, 20, 665, 10.1200/JCO.2002.20.3.665

Gandhi, 2003, Pharmacokinetics and pharmacodynamics of plasma clofarabine and cellular clofarabine triphosphate in patients with acute leukemias, Clin Cancer Res, 9, 6335

Sampath, 2006, Pharmacodynamics of cytarabine alone and in combination with 7-hydroxystaurosporine (UCN-01) in AML blasts in vitro and during a clinical trial, Blood, 107, 2517, 10.1182/blood-2005-08-3351

Cavelier, 2009, Constitutive activation of the DNA damage signaling pathway in acute myeloid leukemia with complex karyotype: potential importance for checkpoint targeting therapy, Cancer Res, 69, 8652, 10.1158/0008-5472.CAN-09-0939

Thông tin
Thông tin xuất bản

Molecular Cancer Therapeutics

Tập 10 Số 4

591-602

Thông tin tác giả