New Succinimides with Potent Anticancer Activity: Synthesis, Activation of Stress Signaling Pathways and Characterization of Apoptosis in Leukemia and Cervical Cancer Cells
Abstract
:1. Introduction
2. Results
2.1. Synthesis
2.2. MTT Cytotoxicity Studies
2.3. Activation of Apoptosis and Expression Profile of Apoptotic Genes
2.4. Effect of Dicarboximides on MAP Kinase Signaling Pathways
2.5. Interaction with DNA
3. Discussion
4. Materials and Methods
4.1. Chemistry
4.1.1. Synthesis of Imides 1–5 and 1h
4.1.2. Synthesis of 1,7-Dimethyl-8,9-diphenyl-4-azatricyclo[5.2.1.02,6]dec-8-ene-3,5,10-trione (1)
4.1.3. Synthesis of 1,7-Diethyl-8,9-tetraphenyl-4-azatricyclo[5.2.1.02,6]dec-8-ene-3,5,10-trione (2)
4.1.4. Synthesis of 1,7,8,9-Tetraphenyl-4-azatricyclo[5.2.1.02,6]dec-8-ene-3,5-dione (3)
4.1.5. Synthesis of N-Hydroxyethylimides 4 and 5
- - 2,5-dimethyl-3,4-diphenylcyclopenta-2,4-dien-1-one (0.02 mol) for compound 4
- - 1,2,3,4-tetraphenylcyclopenta-1,3-diene (0.02 mol) for compound 5 and N-hydroxymaleimide (0.022 mol) was heated for 6 h in 20 mL of benzene. When the reaction was completed, the solvent was evaporated under reduced pressure, and the solid residue was crystallized from benzene.
4.1.6. Synthesis of 4-(2-Chloroethyl)-1,7-dimethyl-8,9-diphenyl-4-azatricyclo[5.2.1.02,6]dec-8-ene-3,5,10-trione (1h)
4.1.7. General Conditions of Synthesis of New N-Alkylamino Derivatives of 1,7-Dimethyl-8,9-diphenyl-4-azatricyclo[5.2.1.02,6]dec-8-ene-3,5,10-trione (1i–1j)
4.1.8. General Conditions of the Synthesis of New Alkanoloamine Derivatives of Dicarboximides (1b–1g, 2b–2g, 3b–3g)
- 1: 1,7-dimethyl-8,9-diphenyl-4-azatricyclo[5.2.1.02,6]dec-8-ene-3,5,10-trione
- 2: 1,7-diethyl-8,9-diphenyl-4-azatricyclo[5.2.1.02,6]dec-8-ene-3,5,10-trione
- 3: 1,7,8,9-tetraphenyl-4-azatricyclo[5.2.1.02,6]dec-8-ene-3,5-dione
4.1.9. Preparation of Acetate Salt of Derivatives Ic, Ie, II
4.1.10. Determination of Lipophilicity by Reversed-Phase Chromatography
4.2. Anticancer Activity
4.2.1. Cells and Cytotoxicity Assay
4.2.2. Analysis of Cell Apoptosis by Caspase-3/7 Activity Assay
4.2.3. Gene Expression Analysis with DNA Microarray
4.2.4. MAPKs Immunoblotting
4.2.5. Digestion of Plasmid DNA with BamHI Restriction Nuclease
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Imide Group | Compound | clogP (Calculated) | Median clogP | logkw (Experimental) | Median logkw |
---|---|---|---|---|---|
2 | Ia | 3.82 | 4.94 | ||
Ib | 4.63 | 5.09 | |||
Ic | 4.95 | 4.27 | 5.02 | 5.05 | |
Id | 4.27 | 5.05 | |||
Ie | 4.06 | 5.17 | |||
1 | 1b | 2.48 | 4.78 | ||
1c | 3.22 | 4.91 | |||
1d | 3.15 | 4.87 | |||
1e | 4.97 | nd | |||
1f | 3.47 | nd | |||
1g | 2.31 | 3.19 | 4.83 | 4.85 | |
4 | 2.69 | nd | |||
1h | 3.84 | nd | |||
1j | 2.88 | nd | |||
1i | 4.04 | nd | |||
2 | 2b | 3.39 | 5.08 | ||
2c | 4.13 | 4.74 | |||
2d | 4.06 | 5.10 | |||
2e | 5.88 | 4.1 | nd | 5.03 | |
2f | 4.38 | nd | |||
2g | 3.22 | 4.98 | |||
3 | II | 5.45 | 6.34 | ||
3 | 3b | 5.02 | 6.31 | ||
3c | 5.76 | 6.06 | |||
3d | 5.69 | 6.28 | |||
3e | 7.51 | 5.69 | nd | 6.28 | |
3f | 6.01 | nd | |||
3g | 4.85 | 6.28 | |||
5 | 5.23 | nd |
Compound | HeLa | K562 | MOLT-4 | HUVEC |
---|---|---|---|---|
1b | 30 (7.5) | 32 (8.3) | 7 (1.8) | 20 (2.4) |
1c | 26 (5.2) | 28 (6.7) | 28 (5) | 23 (3.2) |
1d | 59 (8.9) | 39 (9.4) | 37 (4.4) | 22 (3.3) |
1e | 3.2 (0.4) | 5.8 (1.0) | 8 (0.9) | 30 (4.2) |
1f | 55 (17.6) | 18 (3.8) | 40 (10.8) | 21 (1.6) |
1g | >100 | >100 | >100 | nd |
4 | 61 (14.8) | 105 (15.2) | 71 (14.8) | 56 (4.8) |
1h | nd | nd | 20 (4.7) | 64 (7) |
1j | >100 | >100 | >100 | nd |
1i | 52 (19.2) | 30 (4.1) | 15 (1.2) | 23 (1.8) |
Ic * | 18 (3.1) | 20 (3.4) | 15 (3.2) | 15 (6.3) |
Ie * | 10 (2.6) | 8 (1.7) | 15 (2.3) | 3.5 (0.2) |
2b | 26 (5.6) | 8 (1.5) | 14 (0.9) | 3.4 (0.3) |
2c | 3.2 (0.8) | 4 (0.5) | 2.5 (0.1) | 3.3 (0.2) |
2d | 18 (4.9) | 5.8 (1.2) | 14 (2.6) | 3.6 (0.2) |
2e | 19 (4.5) | 4.1 (0.5) | 3.1 (0.2) | 1.2 (0.06) |
2f | 5.1 (0.7) | 3.9 (0.5) | 3.4 (0.3) | 3.1 (0.3) |
2g | 23 (3.6) | 20 (1.4) | 18 (2.5) | 17 (1.6) |
II * | 20 (2.4) | 16 (4.5) | 18 (5) | 4 (0.5) |
3b | >100 | >100 | >100 | nd |
3c | 2 (0.4) | 2 (0.6) | 2.6 (0.3) | 0.4 (0.03) |
3d | 1.9 (0.6) | 3.2 (0.4) | 2.2 (0.3) | 0.3 (0.02) |
3e | 40 (8.4) | 6 (1.4) | 5.4 (0.5) | 7 (0.6) |
3f | 3 (0.9) | 3 (0.6) | 3 (1) | 0.4 (0.02) |
3g | 120 (16) | 31 (3.6) | 5.2 (0.9) | 17 (1.8) |
5 | 8 (1) | 6.2 (1) | 2.1 (0.1) | 0.7 (0.08) |
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Cieślak, M.; Napiórkowska, M.; Kaźmierczak-Barańska, J.; Królewska-Golińska, K.; Hawrył, A.; Wybrańska, I.; Nawrot, B. New Succinimides with Potent Anticancer Activity: Synthesis, Activation of Stress Signaling Pathways and Characterization of Apoptosis in Leukemia and Cervical Cancer Cells. Int. J. Mol. Sci. 2021, 22, 4318. https://doi.org/10.3390/ijms22094318
Cieślak M, Napiórkowska M, Kaźmierczak-Barańska J, Królewska-Golińska K, Hawrył A, Wybrańska I, Nawrot B. New Succinimides with Potent Anticancer Activity: Synthesis, Activation of Stress Signaling Pathways and Characterization of Apoptosis in Leukemia and Cervical Cancer Cells. International Journal of Molecular Sciences. 2021; 22(9):4318. https://doi.org/10.3390/ijms22094318
Chicago/Turabian StyleCieślak, Marcin, Mariola Napiórkowska, Julia Kaźmierczak-Barańska, Karolina Królewska-Golińska, Anna Hawrył, Iwona Wybrańska, and Barbara Nawrot. 2021. "New Succinimides with Potent Anticancer Activity: Synthesis, Activation of Stress Signaling Pathways and Characterization of Apoptosis in Leukemia and Cervical Cancer Cells" International Journal of Molecular Sciences 22, no. 9: 4318. https://doi.org/10.3390/ijms22094318