New Uracil Analogs with Exocyclic Methylidene Group as Potential Anticancer Agents

Author(s): Angelika Długosz-Pokorska, Joanna Drogosz, Marlena Pięta, Tomasz Janecki, Urszula Krajewska, Marek Mirowski, Anna Janecka*.

Journal Name: Anti-Cancer Agents in Medicinal Chemistry
(Formerly Current Medicinal Chemistry - Anti-Cancer Agents)

Volume 20 , Issue 3 , 2020

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Graphical Abstract:


Abstract:

Background: Hybrid molecules combining uracil skeleton with methylidene exo-cyclic group were designed in the search for novel anticancer drug candidates.

Objective: Two series of racemic 5-methylidenedihydrouracils, either 1,3-disubstituted or 1,3,6-trisubstituted were synthesized and tested for their possible cytotoxic activity against two cancer cell lines (HL-60 and MCF-7) and two healthy cell lines (HUVEC and MCF-10A). The most cytotoxic analogs were re-synthesized as pure enantiomers. The analog designated as U-332 [(R)-3-(4-bromophenyl)-1-ethyl-5-methylidene-6-phenyldihydrouracil], which had a very low IC50 value in HL-60 cell line (0.77μM) and was the most selective towards cancer cells was chosen for further experiments on HL-60 cell line, in order to determine the possible mechanism involved in its antineoplastic action.

Methods: Cytotoxic activities of compound was assessed by the MTT assay. In order to explore the mechanism of U-332 activity, we performed quantitative real-time PCR analysis of p53 and p21 genes. Apoptosis, cell proliferation and DNA damage in HL-60 cells were determined using the flow cytometry. The ability of U-332 to determine GADD45ɑ protein level in HL-60 cells incubated with U-332 was analyzed by ELISA test.

Results: U-332 was shown to generate excessive DNA damage (70% of the cell population), leading to p53 activation, resulting in p21 down-regulation and a significant increase of GADD45α protein, responsible for the cell cycle arrest in G2/M phase.

Conclusion: U-332 can be used as a potential lead compound in the further development of novel uracil analogs as anticancer agents.

Keywords: 5-methylidenedihydrouracils, MTT assay, apoptosis, cell cycle, GADD45 proteins, proliferation, DNA damage.

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Article Details

VOLUME: 20
ISSUE: 3
Year: 2020
Page: [359 - 368]
Pages: 10
DOI: 10.2174/1871520619666191211104128
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