DNA2 Inhibitor C5

DNA2 Inhibitor C5 may be used to evaluate the on-target effects on DNA2 in cancer cells.

DNA2 Inhibitor C5 suppresses nuclease/helicase activities, impairs DNA repair/replication, causes cell cycle arrest, and shows synergistic cancer cell killing with DNA-damaging agents and PARP/ATR inhibitors.

The DNA2 inhibitor C5 (DNA2i-C5) binds to a pocket near the DNA binding site in the helicase domain of DNA2 with an IC50 of 20-30 μM for inhibiting nuclease activity and DNA binding. C5 effectively suppresses both the nuclease and helicase activities of DNA2, impairing DNA end resection, homologous recombination, single-strand annealing repair, and replication fork restart after stalling. In cells, C5 mimics genetic knockout of DNA2, causing cell cycle arrest in late S/G2 phase, reducing mitotic entry, and leading to chromosomal segregation defects, particularly affecting centromeric DNA replication and integrity. Importantly, C5 shows synergistic killing of various cancer cell types when combined with DNA damaging agents like camptothecin or inhibitors of PARP and ATR, highlighting its potential as a cancer therapeutic strategy.