Enhancement of hypoxia-activated prodrug TH-302 anti-tumor activity by Chk1 inhibition

Abstract

BackgroundThe hypoxia-activated prodrug TH-302 is reduced at its nitroimidazole group and selectively under hypoxic conditions releases the DNA cross-linker bromo-isophosphoramide mustard (Br-IPM). Here, we have explored the effect of Chk1 inhibition on TH-302-mediated pharmacological activities.MethodsWe employed in vitro cell viability, DNA damage, cellular signaling assays and the in vivo HT29 human tumor xenograft model to study the effect of Chk1inhibition on TH-302 antitumor activities.ResultsTH-302 cytotoxicity is greatly enhanced by Chk1 inhibition in p53-deficient but not in p53-proficient human cancer cell lines. Chk1 inhibitors reduced TH-302-induced cell cycle arrest via blocking TH-302-induced decrease of phosphorylation of histone H3 and increasing Cdc2-Y15 phosphorylation. Employing the single-cell gel electrophoresis (comet) assay, we observed a potentiation of the TH-302 dependent tail moment. TH-302 induced γH2AX and apoptosis were also increased upon the addition of Chk1 inhibitor. Potentiation of TH-302 cytotoxicity by Chk1 inhibitor was only observed in cell lines proficient in, but not deficient in homology-directed DNA repair. We also show that combination treatment led to lowering of Rad51 expression levels as compared to either agent alone. In vivo data demonstrate that Chk1 inhibitor enhances TH-302 anti-tumor activity in p53 mutant HT-29 human tumor xenografts, supporting the hypothesis that these in vitro results can translate to enhanced in vivo efficacy of the combination.ConclusionsTH-302-mediated in vitro and in vivo anti-tumor activities were greatly enhanced by the addition of Chk1 inhibitors. The preclinical data presented in this study support a new approach for the treatment of p53-deficient hypoxic cancers by combining Chk1 inhibitors with the hypoxia-activated prodrug TH-302.

Engineered Breast Cancer Cell Spheroids Reproduce Biologic Properties of Solid Tumors

“…We note that only the larger (hypoxic) spheroids were co‐treated due to the hypoxia‐dependent mechanism of TH‐302. Consistent with previous studies, TH‐302 alone at the concentrations used was ineffective . However, a major reduction in drug resistance was observed at 100 × 10 −9 m doxorubicin with increasing TH‐302 concentration ( Figure ), suggesting that these compounds synergistically target cancer cells.…”

The Cdc2/Cdk1 inhibitor, purvalanol A, enhances the cytotoxic effects of taxol through Op18/stathmin in non-small cell lung cancer cells in vitro

“…In this study, we demonstrated that the Cdc2/CDK1 inhibitor, purvalanol A, effectively enhanced the sensitivity of NCI-H1299 cells to taxol and inhibited cellular proliferation and colony formation. Other studies have also confirmed that CDK1 inhibition significantly enhances drug-induced colony formation and apoptosis in breast cancer cells and colon carcinoma cells and glioma cells (25)(26)(27).…”

Molecular Pathways: Hypoxia-Activated Prodrugs in Cancer Therapy

“…Combination of TH-302 with Chk1 inhibitor AZD-7762 had greater efficacy in a colorectal xenograft model than either agent alone. This sensitization was shown to be due to disruption of the Chk1-mediated DNA damage checkpoint of the cell cycle and induction of apoptosis, providing additional support to the preclinical translational rationale for combining TH-302 with a Chk1 inhibitor (59). Alternatively, the concept of hypoxia-activatable Chk1 inhibitors was realized in the compound CH-01, a HAP that releases Chk1/Aurora A inhibitor following reduction of a 4-nitrobenzyl in hypoxic conditions (Fig.…”

“…showed that TH-302 cytotoxicity was greatly enhanced by Chk1 inhibition in p53-deficient human cancer cell lines. Chk1 inhibitors reduced TH-302–induced cell cycle arrest by increasing histone H3 and Cdc2-Y15 (59). Combination of TH-302 with Chk1 inhibitor AZD-7762 had greater efficacy in a colorectal xenograft model than either agent alone.…”