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

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.

An Experimental and Theoretical Study of the 1,3-Dipolar Cyclo­addition of Alloxan-Derived Azomethine Ylides to Cyclopropenes

“…The results of these investigations are presented in Figure 2. Based on our previous studies, [20][21][22]24,29 we selected phenyl (4a), carboxymethyl (4e) and N-isopropylcarbamoyl (4f) substituted cyclopropa[a]pyrrolizines along with spirobarbiturate-3-azabicyclo[3.1.0]hexanes containing either a methyl (6b, 6j) or an isobutyl (6g, 6o, 6t) substituent for this preliminary study. Phenyl-substituted cyclopropane ring cycloadducts were found to be more active in both cases (compare 4a vs. 4e or 4f and 6b or 6g vs. 6j, 6o or 6t).…”

11H-Benzo[4,5]imidazo[1,2-a]indol-11-one as a New Precursor of Azomethine Ylides: 1,3-Dipolar Cycloaddition Reactions with Cyclopropenes and Maleimides

“…We used derivatives of isatin [ 22 ], alloxan [ 23 ], ninhydrin [ 24 , 25 , 26 , 27 ], 11 H -indeno[1,2- b ]quinoxalin-11-one [ 28 ], and tryptanthrin [ 29 ] as the ketone components. The products of these reactions were pharmacologically interesting spiro-fused 3-azabicyclo[3.1.0]hexanes and pyrrolizines, some of which were identified as exhibiting in vitro antitumor activity [ 30 , 31 ].…”

Identification of Spiro-Fused Pyrrolo[3,4-a]pyrrolizines and Tryptanthrines as Potential Antitumor Agents: Synthesis and In Vitro Evaluation

“…In our recent works it was shown that (spiro) azabicyclo[3.1.0]hexanes and (spiro) cyclopropa[ a ]pyrrolizines are readily available from cyclopropenes and azomethine ylides (generated in situ from amino acids or peptides and carbonyl compounds such as ninhydrins, indenoquinoxalines, pyrroloisoquinolines and tryptanthrines) and some adducts inhibited the cancer cells grows in vitro [ 33 , 34 , 35 , 36 , 37 ]. In addition, the morphological changes of cells occurring under the action of these compounds (in particular, such cytoskeleton components as actin microfilaments) suggest that the studied substances have not only cytostatic activity, but also can lead to a decrease in metastasis of tumor cells, which opens up broad prospects for the study of their action in vivo [ 38 ].…”