Potent Apoptotic Response Induced by Chloroacetamidine Anthrathiophenediones in Bladder Cancer Cells
Susanna Cogoi1, Sonia Zorzet2, Andrey E Shchekotikhin3
1†Department of Medical and Biological Sciences, University of Udine, P.le Kolbe 4, 33100 Udine, Italy.
Abstract:
We previously found that two neighboring G-quadruplexes behave as a molecular switch controlling the expression of HRAS (Cogoi, S.; Schekotikhin, A. E.; Xodo, L. E. Nucl. Acids Res. 2014, DOI: 10.1093/nar/gku574). In this study we have designed anthrathiophenediones with two chloroacetamidine-containing side chains (CATDs) as G-quadruplex binders and have examined their anticancer activity in T24 bladder cancer cells bearing mutant HRAS and in T24 xenografts. The designed CATDs (3a-e), bearing alkyl side chains of different length, penetrate T24 cancer cells more than their analogues with guanidine-containing side chains. The lead compounds 3a and 3c inhibit HRAS expression, metabolic activity, and colony formation in T24 cancer cells. They also activate a strong apoptotic response, as indicated by PARP-1, caspases 3/7, and annexin V/propidium iodide assays. Apoptosis occurs under conditions where cyclin D1 is down-regulated and the cell cycle arrested in G2 phase. Finally, compound 3a inhibits the growth of T24 xenografts and increases the median survival time of nude mice.
Insights
New G-quadruplex binders, chloroacetamidine-containing diones (CATDs), effectively inhibit HRAS expression and cancer cell growth. Lead compounds demonstrate potent anticancer activity in bladder cancer cells and xenografts, offering a promising therapeutic strategy.
Area of Science:
- Medicinal Chemistry
- Molecular Biology
- Oncology
Background:
- G-quadruplexes regulate HRAS gene expression, acting as a molecular switch.
- Targeting oncogene HRAS is a key strategy in cancer therapy.
Purpose of the Study:
- Design and synthesize novel G-quadruplex binders, chloroacetamidine-containing diones (CATDs).
- Evaluate the anticancer activity of CATDs in T24 bladder cancer cells and xenografts.
- Investigate the molecular mechanisms underlying CATD-induced cancer cell death.
Main Methods:
- Synthesis of anthrathiophenedione derivatives with chloroacetamidine side chains (CATDs).
- In vitro assays: cell viability, colony formation, apoptosis markers (PARP-1, caspases 3/7, Annexin V/PI), cell cycle analysis.
- In vivo studies: T24 xenograft models in nude mice.
Main Results:
- Designed CATDs exhibit enhanced cellular uptake in T24 cells compared to analogues.
- Lead compounds 3a and 3c significantly inhibit HRAS expression, metabolic activity, and colony formation.
- CATDs induce apoptosis, down-regulate cyclin D1, and arrest the cell cycle at G2 phase.
- Compound 3a effectively inhibits tumor growth in vivo and improves median survival in mice.
Conclusions:
- Chloroacetamidine-containing diones are effective G-quadruplex binders with significant anticancer potential.
- Targeting HRAS with CATDs offers a promising therapeutic approach for bladder cancer.
- Compound 3a demonstrates efficacy in preclinical models, warranting further investigation.
