Substitution at the F-ring N-imide of the indolocarbazole antitumor drug NB-506 increases the cytotoxicity, DNA binding, and topoisomerase I inhibition activities.

Abstract

The antitumor drug NB-506, which is currently undergoing phase I/II clinical trials, contains a DNA-intercalating indolocarbazole chromophore substituted with a glucose residue. In addition to interacting with DNA, the drug stabilizes the topoisomerase I-DNA covalent complex. To reinforce the DNA binding and anti-topoisomerase I activities of NB-506, an analogue containing a new substituent on the naphthalimide ring F was synthesized. The N-formylamino group of NB-506 has been replaced with a more hydrophilic group, N-bis(hydroxymethyl)methylamino. In this study we show that the incorporation of a longer substituent on the N6 position effectively reinforces both the interaction with DNA and the capacity of the drug to maintain the integrity of the topoisomerase I-DNA covalent complexes. The strength and the mode of binding of the drugs to DNA were studied by complementary biophysical techniques including absorption, fluorescence, and circular and linear dichroism. Various biochemical procedures were applied to investigate the effects on human topoisomerase I using plasmid DNA as well as restriction fragments. The drug binding sites and the positions of the topoisomerase I-mediated cleavage sites were mapped with nucleotide resolution using footprinting and sequencing techniques. Cytotoxicity measurements performed with various human cancer cell lines (HCT-116, DLD-1, MKN-45) indicate that the newly designed drug is 3 to 4 times more toxic to colon and gastric cancer cells than NB-506. Therefore, the results suggest that the antitumor activity of indolocarbazole-based drugs can be enhanced by incorporating DNA and/or topoisomerase I reactive groups. They also support the hypothesis that the substituent on the imide nitrogen on the F ring of NB-506 has direct interaction with the molecular target. The study helps to define the structure-activity relationships in the indolocarbazole series of antitumor agents targeting topoisomerase I.

• Authors: C Bailly, X Qu, J B Chaires, P Colson, C Houssier, M Ohkubo, S Nishimura, T Yoshinari
• Journal: Journal of medicinal chemistry (J Med Chem) Vol. 42 Issue 15 Pg. 2927-35 (Jul 29 1999) ISSN: 0022-2623 [Print] United States
• PMID: 10425102 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Antineoplastic Agents
• Carbazoles
• DNA, Neoplasm
• Enzyme Inhibitors
• Glucosides
• Intercalating Agents
• NB 506
• Topoisomerase I Inhibitors
• DNA

Topics

• Animals
• Antineoplastic Agents (chemical synthesis, chemistry, metabolism, pharmacology)
• Carbazoles (chemical synthesis, chemistry, metabolism, pharmacology)
• Cattle
• Circular Dichroism
• DNA (metabolism)
• DNA, Neoplasm (metabolism)
• Drug Screening Assays, Antitumor
• Enzyme Inhibitors (chemical synthesis, chemistry, metabolism, pharmacology)
• Glucosides (chemical synthesis, chemistry, metabolism, pharmacology)
• Humans
• Inhibitory Concentration 50
• Intercalating Agents (chemical synthesis, chemistry, metabolism, pharmacology)
• Sequence Analysis, DNA
• Spectrometry, Fluorescence
• Spectrophotometry, Ultraviolet
• Structure-Activity Relationship
• Topoisomerase I Inhibitors
• Tumor Cells, Cultured