Abstract 3957: Anticancer effect of novel 1,4-benzodiazepine derivatives through tubulin polymerization inhibition

Abstract

Abstract Benzodiazepines are well known as a key scaffold in medicinal chemistry, providing bioactive compounds for a wide variety of biological targets. Among their primary use as anxiolytics and sedatives, the 1,4-benzodiazepine derivatives and their metabolites play a role as antitumor, antithrombotic, antiviral and antimalarial agents. Recent studies revealed the influence of the benzodiazepine derivatives on microtubule assembly or stability with subsequent mitotic failure. These microtubule-targeting effects were evaluated in novel benzodiazepine derivatives. The antiproliferative activities of novel derivatives were tested against a panel of 10 cancer cell lines and 2 non-cancer cell lines. The compounds with IC50<10 µM were further analyzed for the cell cycle. Flow cytometry and fluorescent cell cycle indicator (FUCCI) assays were performed to evaluate the effects of derivatives on the cell cycle phases. The immunofluorescence alpha-tubulin assay and tubulin polymerization assay were accomplished to detect the changes in the mitosis. Implementation of molecular docking revealed the specific binding site of the derivatives. All tested derivatives reveal the G2/M block in the cell cycle and 3 of them were identified as M-phase blockers. The immunofluorescence alpha-tubulin assay detected the disruption of the cellular microtubule network, and the tubulin polymerization assay uncovered the potential of the tested compounds to inhibit tubulin polymerization. Molecular docking confirms that benzodiazepine derivatives fit the colchicine site best and are partially overlaid with the native colchicine binding pose. This suggests that these compounds may adapt to the tubulin binding site in a conformation induced by colchicine The inhibition of tubulin polymerization by novel derivatives is one of their possible mechanisms of action on antiproliferative effect on cancer cell lines. More rigorous studies will be performed further to establish pose stability and explain structure-activity relationships. This work was supported by European Union - Programme EXCELES, ID Project No. LX22NPO5102, the Czech Ministry of Education, Youth and Sports (CZ-OPENSCREEN - LM2018130, EATRIS-CZ - LM2018133), and by the internal grant of Palacky University Olomouc (IGA_LF_2022_033) and European Regional Development Fund - Project ENOCH (No. CZ.02.1.01/0.0/0.0/16_019/0000868). Citation Format: Alzbeta Srovnalova, Jiri Rehulka, Sona Gurska, Pavel Polyshchuk, Sergey Bachinsky, Petr Dzubak, Marian Hajduch. Anticancer effect of novel 1,4-benzodiazepine derivatives through tubulin polymerization inhibition. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3957.