Dados do Trabalho

Título

Evaluation of Antitumor Activity and In vitro Toxicity of Hybrid Compounds (2-Arylquinazolinochalcones) against Breast Cancer Cell Lines

Introdução

The development of antineoplastic drugs hinges upon uncovering efficient and precise molecules that target neoplastic cells. In this context, hybrid molecules have been highlighted in recent years due to their synergistic effects at reduced dosages and lower probability of resistance development, in contrast to their individual applications in therapies. Chalcones and quinazolines emerge as potential candidates, given their capacity to inhibit factors linked to diverse cellular functions, such as survival and resistance, both in their isolated and conjugated forms.

Objetivo

To assess the antineoplastic potential of novel hybrid molecules of chalcones and quinazolines in vitro, as well as their in silico pharmacokinetic, physicochemical, and molecular target parameters.

Métodos

We evaluated the cytotoxic potential of fifteen 2-Arylquinazolinochalcones molecules (R1 – R15) and determined their IC50 values through the Sulforhodamine B assay, adapted from the One and Five Dose Screening protocols of the National Cancer Institute (NCI). DMSO and 5-Fluorouracil (5-FU) were used as negative and positive controls, respectively. Based on the IC50 values, we calculated the compounds selectivity indexes by using IC50 values of non-tumoral cell lines (HFF-1 and HaCat). Additionally, we examined cell death and cell cycle parameters in the triple-negative breast tumor cell line (MDA-MB-231), noted for its heightened sensitivity, using flow cytometry. Evaluations of molecular changes in proliferative signaling pathways were conducted via phospho-MAPK protein arrays (R&D Systems). The hybrid molecules were also assessed for their physicochemical and pharmacokinetic parameters, as well as their molecular target prediction, using in silico platforms (ADMETLab and SwissADME).

Resultados

Following initial screening, molecules R2 and R14 exhibited higher selectivity for breast cancer cell lines (IS>2) of three different molecular subtypes at a dosage of 10μM, using non-tumoral cell lines HFF-1 and HaCat as selectivity parameters. Both molecules R2 and R14 showed IC50 values below 5μM for MDA-MB-231 (2.93 and 2.45 μM) and T-47D (4.07 and 1.21 μM) tumor cell lines, respectively. Moreover, these molecules increased the proportion of cells in early and/or late-stage apoptosis at 48 and 72 hours of treatment, and molecule R2 contributed to heightened cell retention in the G0/G1 cell cycle phase, as evidenced by reduced thymidine kinase expression post-treatment. The MAPK protein expression profile indicated a reduction in phosphorylated AKT levels following R2 treatment, compared to the negative control. In silico analyses suggested that both molecules are potential inhibitors of ABCG2 and Pgp-1 proteins, and they also exhibited favorable parameters for absorption and distribution, with low probabilities of blood-brain barrier penetration.

Conclusões

R2 and R14 exhibit targeted antitumor activity against breast cancer cells, with superior efficacy when compared to 5-FU treatment, and they led to increased cell death, impaired cell cycle progression, and inhibition of AKT phosphorylation. These molecules also display favorable pharmacokinetic parameters and are potential inhibitors of proteins linked to resistance development. A subsequent evaluation will be conducted using 3D culture models.

Palavras-chave

chalcones; quinazolines; hybrid drugs.

Financiador do resumo

CAPES, FAPESP, PAIP (Barretos Cancer Hospital), CNPq, FAPERGS.