Inhibitors Against Chk1 And Wee1, Mk-8776 And Mk-1775, Strongly Synergize In Vitro And In Vivo To Inhibit Tumor Growth

Inhibitors of the checkpoint kinases CHK1 and WEE1 are under development as sensitizers to chemotherapy or radiation therapy. Disruption of either CHK1 or WEE1 activity compromises the cell cycle arrest that normally accompanies the DNA damage response, forcing cells to prematurely divide before repairing damaged DNA. However, inhibition of either CHK1 or WEE1 alone also disrupts DNA replication, resulting in DNA double strand breaks (DSBs) primarily in S-phase cells. Here we describe the ability of inhibitors against CHK1 and WEE1 to synergize both in vitro and in vivo. MK-8776 and MK-1775 are advancing through early clinical trials as potent and selective inhibitors of CHK1 and WEE1, respectively. MK-8776 increases the cell-based potency of MK-1775 in multiple cancer cell lines by as much as a 10-fold EC50 shift in proliferation assays. Staining for the DSB marker γH2AX at low concentrations of each drug demonstrates that MK-8776 and MK-1775 in combination induce up to 50-fold more DNA damage than either MK-8776 or MK-1775 alone. The ability of the combination to drive premature mitosis, determined by pHH3 quantitation, was variable, suggesting that generation of DSBs rather than G2/M checkpoint abrogation is the mechanism underlying combination cytotoxicity. Induction of DSBs requires cyclin-dependent kinase activity and takes place largely in S-phase cells, implying deregulation of DNA replication. Furthermore, staining for DNA damage response markers γH2AX and pCHK1-ser345 was both more intense and more durable in a LoVo xenograft model of cancer when MK-8776 and MK-1775 were dosed together compared to either compound alone. At tolerated doses, the combination demonstrated significantly greater anti-tumor efficacy than either MK-8776 or MK-1775 alone in LoVo xenografts (91% tumor growth inhibition for the combination versus 28% and 41% for MK-8776 and MK-1775, respectively). Similar observations were made in additional xenograft models. These results demonstrate in vitro and in vivo synergy between MK-8776 and MK-1775, indicating unique and complimentary anti-tumor effects of CHK1 and WEE1 inhibition. Our studies provide preclinical rationale for combining WEE1 and CHK1 inhibitors as a novel cancer treatment regimen. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 2051. doi:1538-7445.AM2012-2051