Cell cycle G(2) checkpoint abrogation is an attractive strategy for sensitizing
cancer cells to
DNA-damaging
anticancer agent without increasing adverse effects on normal cells. However, there is no single proven molecular target for this therapeutic approach. High-throughput screening for molecules inhibiting CHK1, a
kinase that is essential for the G(2) checkpoint, has not yet yielded therapeutic G(2) checkpoint inhibitors, and the
tumor suppressor phenotypes of ATM and CHK2 suggest they may not be ideal targets. Here, we optimized two G(2) checkpoint-abrogating
peptides, TAT-S216 and TAT-S216A, based on their ability to reduce G(2) phase accumulation of
DNA-damaged cells without affecting M phase accumulation of cells treated with a microtubule-disrupting compound. This approach yielded a
peptide CBP501, which has a unique, focused activity against molecules that phosphorylate Ser(216) of CDC25C, including MAPKAP-K2, C-Tak1, and CHK1.
CBP501 is >100-fold more potent than TAT-S216A and retains its selectivity for
cancer cells.
CBP501 is unusually stable, enters cells rapidly, and increases the cytotoxicity of
DNA-damaging anticancer drugs against
cancer cells without increasing adverse effects. These findings highlight the potency of
CBP501 as a G(2)-abrogating
drug candidate. This report also shows the usefulness of the cell cycle phenotype-based protocol for identifying G(2) checkpoint-abrogating compounds as well as the potential of
peptide-based compounds as focused multitarget inhibitors.