HU-331 is a catalytic inhibitor of topoisomerase IIα.

Abstract

Topoisomerases are essential enzymes that are involved in DNA metabolism. Topoisomerase II generates transient DNA strand breaks that are stabilized by anticancer drugs, such as doxorubicin, causing an accumulation of DNA damage. However, doxorubicin causes cardiac toxicity and, like etoposide and other topoisomerase II-targeted agents, can induce DNA damage, resulting in secondary cancers. The cannabinoid quinone HU-331 has been identified as a potential anticancer drug that demonstrates more potency in cancer cells with less off-target toxicity than that of doxorubicin. Reports indicate that HU-331 does not promote cell death via apoptosis, cell cycle arrest, caspase activation, or DNA strand breaks. However, the precise mechanism of action is poorly understood. We employed biochemical assays to study the mechanism of action of HU-331 against purified topoisomerase IIα. These assays examined DNA binding, cleavage, ligation, relaxation, and ATPase activities of topoisomerase IIα. Our results demonstrate that HU-331 inhibits topoisomerase IIα-mediated DNA relaxation at micromolar levels. We find that HU-331 does not induce DNA strand breaks in vitro. When added prior to the DNA substrate, HU-331 blocks DNA cleavage and relaxation activities of topoisomerase IIα in a redox-sensitive manner. The action of HU-331 can be blocked, but not reversed, by the presence of dithiothreitol. Our results also show that HU-331 inhibits the ATPase activity of topoisomerase IIα using a noncompetitive mechanism. Preliminary binding studies also indicate that HU-331 decreases the ability of topoisomerase IIα to bind DNA. In summary, HU-331 inhibits relaxation activity without poisoning DNA cleavage. This action is sensitive to reducing agents and appears to involve noncompetitive inhibition of the ATPase activity and possibly inhibition of DNA binding. These studies provide a promising foundation for the exploration of HU-331 as a catalytic inhibitor of topoisomerase IIα.

• Authors: Kellie M Regal, Susan L Mercer, Joseph E Deweese
• Journal: Chemical research in toxicology (Chem Res Toxicol) Vol. 27 Issue 12 Pg. 2044-51 (Dec 15 2014) ISSN: 1520-5010 [Electronic] United States
• PMID: 25409338 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antigens, Neoplasm
• DNA-Binding Proteins
• Topoisomerase II Inhibitors
• cannabidiol hydroxyquinone
• Cannabidiol
• DNA
• DNA Topoisomerases, Type II

Topics

• Antigens, Neoplasm (metabolism)
• Cannabidiol (analogs & derivatives, pharmacology)
• Catalysis
• DNA (metabolism)
• DNA Topoisomerases, Type II (metabolism)
• DNA-Binding Proteins (antagonists & inhibitors, metabolism)
• Topoisomerase II Inhibitors (pharmacology)