Mitomycin C (MC) is a naturally occurring alkylating agent used as an anticancer drug for its ability to form toxic DNA interstrand cross-links (ICLs). Aziridinomitosenes (AZMs) are structurally similar to MC and were discovered as metabolites of MC. AZMs display similar anticancer properties as MC through the formation of DNA-ICLs, however it has been observed that AZMs with unsubstituted C6/C7 positions can form DNA-ICLs in non-reductive conditions, something not previously observed by MC. Additionally, AZMs have been shown to form DNA-protein cross-links (DPCs). To better understand the role of C6/C7 sites in the mechanism of ICL formation, four AZM analogs with varying methylated and unsubstituted C6 and C7 sites are being synthesized. It is expected that analogs with substituted C6/C7 positions will generate less DPC/ICLs compared to analogs with unsubstituted C6/C7 positions. To test this hypothesis, various biochemical assays will be used to assess each analog’s effects on cell proliferation, DNA-ICL formation, and DPC formation.