Dados do Trabalho

Título

Preclinical evaluation of antitumor activity and toxicity of novel PI3K inhibitors in colorectal tumor cell lines

Introdução

In oncogenesis, the deregulation of the phosphatidylinositol-3-kinase (PI3K) pathway is associated with a wide type of cancers, especially colon and rectum. As a result, PI3K inhibitors have emerged as a promising target for antineoplastic therapy. Recent advances in PI3K-α (PIK3CA) hyperactivity inhibition have shown a reduction in tumor progression. In this study, we aimed to assess the specificity and inhibitory activity of 10 novel unreported PI3K inhibitors in colorectal tumor cell lines.

Objetivo

To assess the antineoplastic potential of recently synthesized novel PI3K inhibitors in colorectal tumor cells using in vitro models.

Métodos

To assess the efficacy of the PI3K inhibitors, the HCT-116, HCT-15, and DIFI colorectal tumor cell lines and the HaCat healthy control cell line were used through the screening protocol adapted from the National Cancer Institute (NCI), consisting of two steps. For the one-dose step, the cell lines were exposed to the 10 inhibitors and to Alpesilib as a positive control at a concentration of 10µM. The five-dose step used the best inhibitors selected in one-dose to determine the IC50 by MTS. Then, a 3D cell culture model of the HCT-166 line was performed to determine the morphological changes of the spheroids and cell viability using Calcein and Propidium Iodide dyes. To evaluate the cell apoptosis, the HCT-116 cell line was treated with the best inhibitors and analyzed by flow cytometry after 72 hours. An in silico analysis was performed using the ADMETlab prediction platform to evaluate physicochemical, pharmacokinetic, and toxicity parameters.

Resultados

After 72 hours of treatment in the one-dose and five-dose steps, a reduction in the cell growth of tumor cell lines (<50%) was observed for the compounds VL311 and VL336. The IC50 for VL311 were 1.59µM, 3.84µM, 92.77µM and 2.85µM for HCT-116, HCT-15, DIFI, and HaCat respectively. The IC50 for VL336 were 1.51µM, 2.83µM, 6.03µM and 3.1µM. Both compounds presented lower IC50 when compared to Alpesilib, which were 12.18µM, 165µM, 13.17µM, and 7.55µM. The 3D model revealed a reduction of spheroids’ perimeter associated with lower cell viability observed under the fluorescence assay when compared with Alpelisib. The flow cytometry assay revealed that VL311 has shown to induce 10% of early apoptosis and 20% of late apoptosis, while VL336 induced 7% and 15%, and Alpelisib induced 10% and 25%. The in silico analysis revealed that both compounds exhibit moderately hydrophobic and medium to high lipophilicity properties, hepatotoxicity, and ocular sensitivity.

Conclusões

The findings of the present study highlight the potential efficacy of VL336 and VL311 compounds as promising candidates in the treatment of colorectal tumors, requiring future functional studies and in vivo validations for the development of new innovative therapies for colorectal cancer.

Palavras-chave

PI3K inhibitors, preclinical trial, colorectal cancer.

Financiador do resumo

CAPES, PAIP (Barretos Cancer Hospital), MPT (Public Ministry of Labour).