Aziridinomitosenes are synthetic chemotherapeutic agents that are analogs of Mitomycin C, as well as other mitomycins. They are known for their ability to form DNA interstrand cross-links (ICLs), and more recently have been observed to form DNA-protein cross-links (DPCs). In this work, three synthetic aziridinomitsenes were prepared and studied to better understand the role of the C6 and C7 positions in the formation of DNA-protein cross-links. In addition, two separate methods, the advanced recovery of K-SDS precipitates and the K-SDS assays were utilized and compared to assess the advancements in the analysis of DPC formation. H AZM, C6 AZM, Dimethyl AZM, and Mitomycin C were compared to formaldehyde, a known DNA-protein cross-linking agent in these techniques. Results from the ARK assay displayed a 4-fold increase in average response in nearly every treatment group when compared to the previously developed K-SDS method. Additionally, the ARK assay provided tighter standard deviations indicating more reliable data. Analysis of ARK data displayed that the C6 AZM was capable of forming a greater percentage of DNA-protein cross-links compared to the H AZM and Dimethyl AZM. These results indicated that the C6 and C7 positions of aziridinomitosenes play an important role in DNA-protein cross-link formation
