Identification of Spiro-Fused [3-azabicyclo[3.1.0]hexane]oxindoles as Potential Antitumor Agents: Initial In Vitro Evaluation of Anti-Proliferative Effect and Actin Cytoskeleton Transformation in 3T3 and 3T3-SV40 Fibroblast.

Nickolay A Knyazev,Alexander S Filatov,Sofya A Pechkovskaya,Natalia A Filatova,Stanislav V Shmakov,Alexander V Stepakov,Vitali M Boitsov

doi:10.3390/ijms22158264

Abstract

Novel heterocyclic compounds containing 3-spiro[3-azabicyclo[3.1.0]hexane]oxindole framework (4a, 4b and 4c) have been studied as potential antitumor agents. The in silico ADMET (adsorption, distribution, metabolism, excretion and toxicity) analysis was performed on 4a–c compounds with promising antiproliferative activity, previously synthetized and screened against human erythroleukemic cell line K562 tumor cell line. Cytotoxicity of 4a–c against murine fibroblast 3T3 and SV-40 transformed murine fibroblast 3T3-SV40 cell lines were evaluated. The 4a and 4c compounds were cytotoxic against 3T3-SV40 cells in comparison with those of 3T3. In agreement with the DNA cytometry studies, the tested compounds have achieved significant cell-cycle perturbation with higher accumulation of cells in G0/G1 phase. Using confocal microscopy, we found that with 4a and 4c treatment of 3T3 cells, actin filaments disappeared, and granular actin was distributed diffusely in the cytoplasm in 82–97% of cells. The number of 3T3-SV40 cells with stress fibers increased to 7–30% against 2% in control. We discovered that transformed 3T3-SV40 cells after treatment with compounds 4a and 4c significantly reduced the number of cells with filopodium-like membrane protrusions (from 86 % in control cells to 6–18% after treatment), which indirectly suggests a decrease in cell motility. We can conclude that the studied compounds 4a and 4c have a cytostatic effect, which can lead to a decrease in the number of filopodium-like membrane protrusions.

Highlights

Cancer is one of the fatal diseases, which is characterized by uncontrolled cell division leading to metastasis and invasion into adjacent tissues
Poor absorption or permeation is more likely when there are more than five hydrogen bond donors, ten hydrogen bond acceptors, the molecular weight is greater than 500 g/mol and the calculated log p is greater than 5
The main difference in the sensitivity of 3T3-SV40 cells from 3T3 cells to the effect of the studied compounds was the absence of a change in the number of cells in the G2/M phase relative to the control and only a slight decrease in their number when impacted by cisplatin. These results suggest that compounds 4a and 4c arrest 3T3 and 3T3-SV40 cells at the G0/G1 stage of the cell cycle

Summary

Introduction

Cancer is one of the fatal diseases, which is characterized by uncontrolled cell division leading to metastasis and invasion into adjacent tissues. Synthesizing new anticancer agents with less side effects and increased selectivity is still a developing area of interest in medicinal chemistry [1]. A significant portion of the improved clinical outcomes in cancer can be attributed to refined chemotherapeutic agents. The identification of antitumor natural products and natural product analogs such as Paclitaxel, Vinblastine, Doxorubicin, and Hycamtin has clearly demonstrated the importance of natural products in modern medicine [2]. Many potential drugs fail to reach the clinic because of in silico prediction liabilities. Adsorption, distribution, metabolism, excretion and toxicity (ADMET) processes play a pivotal role in defining the therapeutic efficacy of drugs. Drug likeness appears as a promising paradigm of a compound that optimizes their ADME in the human body [3]

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