Synthesis of N,N-Dimethylcyclohexylamine-Hyptolide and Activity Assessment Against Breast Cancer Stem Cells (BCSCs) Using In Vitro and In Silico Approaches
DOI:
https://doi.org/10.48048/tis.2026.11659Keywords:
Hyptolide, Diels-Alder synthesis, Breast cancer stem cells, Cell cycle arrest, Molecular dockingAbstract
The overexpression of histone deacetylase (HDAC) proteins in breast cancer stem cells (BCSCs) significantly contributes to tumor progression, chemotherapy resistance, and impaired apoptotic signaling through the silencing of tumor suppressor genes. Targeting HDAC with small-molecule inhibitors is therefore considered a promising therapeutic strategy. In this study, a novel hyptolide-based compound, N,N-dimethylcyclohexylamine-hyptolide (compound 3), was successfully synthesized via a Diels-Alder reaction. The compound was structurally characterized using FTIR, 1H-NMR, 13C-NMR and TLC-MS, confirming the formation of a cyclohexene ring bearing a tertiary amine group. The product exhibited a melting point of 83 - 84 °C and a molecular weight of 595 g/mol. In vitro cytotoxicity assays on BCSCs revealed an IC50 value of 47.42 µg/mL, classifying it as moderately active and indicating its potential as a therapeutic candidate for BCSCs. Moreover, flow cytometry analysis demonstrated that the compound induced cell cycle arrest at both the S phase (DNA synthesis) and the G2/M phase (cell division). Complementary in silico molecular docking simulations showed stable binding interactions between the compound and key active-site residues of HDAC, supporting its potential as an HDAC inhibitor. These findings suggest that N,N-dimethylcyclohexylamine-hyptolide may serve as a promising lead for the development of HDAC-targeted therapies, particularly in the treatment of aggressive and drug-resistant breast cancers.
HIGHLIGHTS
- N,N-dimethylcyclohexylamine-hyptolide, as a derivative compound of hyptolide was successfully synthesized via Diels-Alder reaction.
- N,N-dimethylcyclohexylamine-hyptolide exhibits anticancer activity against breast cancer stem cells (BCSCs) by inducing cell cycle arrest in the S and G2/M phases.
- N,N-dimethylcyclohexylamine-hyptolide has potential activity as an HDAC inhibitor based on in silico molecular docking study.
GRAPHICAL ABSTRACT
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