Pixantrone dimaleate is a novel and potent aza-anthracenedione analog that has little cardiotoxicity and anticancer activity. It was formerly known as BBR 2778. It functions as a DNA intercalator and weak inhibitor of topoisomerase II, selectively forming stable DNA adducts at sites of hypermethylation through alkylation. It enters DNA and causes DNA strand crosslinks mediated by topoisomerase II, which inhibits DNA replication and reduces the cytotoxicity of tumor cells. Important oncotherapeutics, anthracenenes and anthracenecyclines are linked to cumulative and irreversible cardiotoxicity when used. In order to minimize treatment-related cardiotoxicity without sacrificing effectiveness, pixantrone was created. Patients diagnosed with aggressive non-Hodgkin lymphoma (aNHL) can benefit from a less cardiotoxic and more effective alternative to doxorubicin: pixantrone. Pixantrone, with IC50s of 37.3 nM, 126 nM, and 136 nM for T47D, MCF-10A, and OVCAR5 cells, respectively, causes cell death in a number of cancer cell lines without disrupting the cell cycle. Pixantrone damages DNA at high concentrations (500 nM), but not at low enough concentrations (100 nM) to cause PANC1 cells to perish. In PANC1 cells, Pixantrone (25 or 100 nM) causes severe chromosomal abnormalities and a mitotic disaster. Because Pixantrone (100 nM) generates merotelic kinetochore attachments that result in chromosome non-disjunction, it may interfere with chromosome segregation. Pixantrone, with IC50s of 0.10 μM, 0.56 μM, 0.058 μM, and 4.5 μM, respectively, potently inhibits the growth of human leukemia K562 cells, etoposide-resistant K/VP.5 cells, MDCK, and ABCB1-transfected MDCK/MDR cells. By acting on topoisomerase IIα, Pixantrone (0.01-0.2 μM) causes a concentration-dependent formation of linear DNA. In an enzymatic reducing system, pyrantrone generates semiquinone free radicals; however, it does not do so in a cellular system, probably because of insufficient cellular absorption. Pixantrone (0.01-10 μM) exhibits strong inhibitory effects on the proliferation of T cells that are specific to the rat 97-116 peptide.