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Characterization and preclinical development of LY2603618: a selective and potent Chk1 inhibitor

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Summary

Interference with DNA damage checkpoints has been demonstrated preclinically to be a highly effective means of increasing the cytotoxicity of a number of DNA-damaging cancer therapies. Cell cycle arrest at these checkpoints protects injured cells from apoptotic cell death until DNA damage can be repaired. In the absence of functioning DNA damage checkpoints, cells with damaged DNA may proceed into premature mitosis followed by cell death. A key protein kinase involved in activating and maintaining the S and G2/M checkpoints is Chk1. Pharmacological inhibition of Chk1 in the absence of p53 functionality leads to abrogation of DNA damage checkpoints and has been shown preclinically to enhance the activity of many standard of care chemotherapeutic agents. LY2603618 is a potent and selective small molecule inhibitor of Chk1 protein kinase activity in vitro (IC50 = 7 nM) and the first selective Chk1 inhibitor to enter clinical cancer trials. Treatment of cells with LY2603618 produced a cellular phenotype similar to that reported for depletion of Chk1 by RNAi. Inhibition of intracellular Chk1 by LY2603618 results in impaired DNA synthesis, elevated H2A.X phosphorylation indicative of DNA damage and premature entry into mitosis. When HeLa cells were exposed to doxorubicin to induce a G2/M checkpoint arrest, subsequent treatment with LY2603618 released the checkpoint, resulting in cells entering into metaphase with poorly condensed chromosomes. Consistent with abrogation of the Chk1 and p53-dependent G2/M checkpoint, mutant TP53 HT-29 colon cancer cells were more sensitive to gemcitabine when also treated with LY2603618, while wild-type TP53 HCT116 cells were not sensitized by LY2603618 to gemcitabine. Treatment of Calu-6 human mutant TP53 lung cancer cell xenografts with gemcitabine resulted in a stimulation of Chk1 kinase activity that was inhibited by co-administration of LY2603618. By all criteria, LY2603618 is a highly effective inhibitor of multiple aspects of Chk1 biology.

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Acknowledgements

We would like to recognize the essential contributions of the Chk1 biology and medicinal chemistry team members from Icos Pharmaceuticals and Array BioPharma: Phyllis Goldman, Laurence Burgess, Erik Christenson, Darcey Clark, Adam Cook, Scott Cowen, Jeff Dantzler, Frank Diaz, Heather Douanpanya, Francine Farouz, Kimba Fischer, John Gaudino, Ryan Holcomb, Angela Judkins, Adam Kashishian, Ed Kesicki, Kim McCaw, Harch Ooi, Vanessa Rada, Fuqiang Ruan, Alex Rudolph, Frank Stappenbeck, Janelle Taylor, Gene Thorsett, Jen Treiberg, Margaret Weidner and Steve White. We would also like to thank Eric Westin and Aimee Bence for constant intellectual input.

Conflict of interests

The authors, Constance King, Henry Diaz, Darlene Barnard, David Barda, David Clawson, Wayne Blosser, Karen Cox, Sherry Guo and Mark Marshall are all employees of the Eli Lilly Company, which supports the development of LY2603618.

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Authors and Affiliations

  1. Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN, 46285, USA

    Constance King, Henry Diaz, Darlene Barnard, David Barda, David Clawson, Wayne Blosser, Karen Cox, Sherry Guo & Mark Marshall

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  1. Constance King
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  2. Henry Diaz
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Correspondence to Mark Marshall.

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King, C., Diaz, H., Barnard, D. et al. Characterization and preclinical development of LY2603618: a selective and potent Chk1 inhibitor. Invest New Drugs 32, 213–226 (2014). https://doi.org/10.1007/s10637-013-0036-7

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