Colchicine Binding Site Agent DJ95 Overcomes Drug Resistance and Exhibits Antitumor Efficacy.

Abstract

Interfering with microtubule dynamics is a well-established strategy in cancer treatment; however, many microtubule-targeting agents are associated with drug resistance and adverse effects. Substantial evidence points to ATP-binding cassette (ABC) transporters as critical players in the development of resistance. Herein, we demonstrate the efficacy of DJ95 (2-(1H-indol-6-yl)-4-(3,4,5-trimethoxyphenyl)-1H-imidazo[4,5-c]pyridine), a novel tubulin inhibitor, in a variety of cancer cell lines, including malignant melanomas, drug-selected resistant cell lines, specific ABC transporter-overexpressing cell lines, and the National Cancer Institute 60 cell line panel. DJ95 treatment inhibited cancer cell migration, caused morphologic changes to the microtubule network foundation, and severely disrupted mitotic spindle formation of mitotic cells. The high-resolution crystal structure of DJ95 in complex with tubulin protein and the detailed molecular interactions confirmed its direct binding to the colchicine site. In vitro pharmacological screening of DJ95 using SafetyScreen44 (Eurofins Cerep-Panlabs) revealed no significant off-target interactions, and pharmacokinetic analysis showed that DJ95 was maintained at therapeutically relevant plasma concentrations for up to 24 hours in mice. In an A375 xenograft model in nude mice, DJ95 inhibited tumor growth and disrupted tumor vasculature in xenograft tumors. These results demonstrate that DJ95 is potent against a variety of cell lines, demonstrated greater potency to ABC transporter-overexpressing cell lines than existing tubulin inhibitors, directly targets the colchicine binding domain, exhibits significant antitumor efficacy, and demonstrates vascular-disrupting properties. Collectively, these data suggest that DJ95 has great potential as a cancer therapeutic, particularly for multidrug resistance phenotypes, and warrants further development. SIGNIFICANCE STATEMENT: Paclitaxel is a widely used tubulin inhibitor for cancer therapy, but its clinical efficacy is often limited by the development of multidrug resistance. In this study, we reported the preclinical characterization of a new tubulin inhibitor DJ95, and demonstrated its abilities to overcome paclitaxel resistance, disrupt tumor vasculature, and exhibit significant antitumor efficacy.

• Authors: Kinsie E Arnst, Yuxi Wang, Zi-Ning Lei, Dong-Jin Hwang, Gyanendra Kumar, Dejian Ma, Deanna N Parke, Qiang Chen, Jinliang Yang, Stephen W White, Tiffany N Seagroves, Zhe-Sheng Chen, Duane D Miller, Wei Li
• Journal: Molecular pharmacology (Mol Pharmacol) Vol. 96 Issue 1 Pg. 73-89 (07 2019) ISSN: 1521-0111 [Electronic] United States
• PMID: 31043459 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2019 by The American Society for Pharmacology and Experimental Therapeutics.

Chemical References

• ATP-Binding Cassette Transporters
• Imidazoles
• Pyridines
• Small Molecule Libraries
• Tubulin
• Tubulin Modulators
• Colchicine

Topics

• ATP-Binding Cassette Transporters (metabolism)
• Animals
• Cell Line, Tumor
• Cell Movement (drug effects)
• Cell Survival (drug effects)
• Colchicine (metabolism)
• Crystallography, X-Ray
• Drug Resistance, Neoplasm (drug effects)
• Female
• HEK293 Cells
• Human Umbilical Vein Endothelial Cells
• Humans
• Imidazoles (administration & dosage, chemistry, pharmacology)
• Male
• Melanoma (drug therapy, metabolism)
• Mice
• Mice, Nude
• Pyridines (administration & dosage, chemistry, pharmacology)
• Small Molecule Libraries (administration & dosage, chemistry, pharmacology)
• Tubulin (chemistry, metabolism)
• Tubulin Modulators (administration & dosage, chemistry, pharmacology)
• Xenograft Model Antitumor Assays