Fig. 5: DNA cleavage assay for the G-A cross-linking agent FGX8-46 (6), and compounds 7 and 8, on the 135 base-pair WnGWn DNA fragment that contains isolated guanines within long tracts of adenines and thymines.
From: DNA sequence-selective G-A cross-linking ADC payloads for use in solid tumour therapies
The WnGWn was incubated with 10, 3, 1, 0.3, 0.1 and 0.3 μM of 6, 7 and 8 (indicated at tops of gels) in 10 mM Tris-HCl, 10 mM NaCl at pH 7.5 overnight at 37 °C, and a dose-dependent effect is evident (Left). The ‘GA’ lanes contain Maxam-Gilbert markers specific for purines, and the ‘con’ lanes are controls in the absence of 6, 7 and 8. The black, red and green arrows on the gels show the locations of cleavage points due to the CBI component of the compounds, thus indicating covalent binding sites. These cleavage sites are also indicated on the DNA sequence using coloured blocks (Bottom Right).
