Tropinone-Derived Alkaloids as Potent Anticancer Agents: Synthesis, Tyrosinase Inhibition, Mechanism of Action, DFT Calculation, and Molecular Docking Studies
Abstract
:1. Introduction
2. Results and Discussion
2.1. Chemical Synthesis and Chemoinformatics Data
2.2. Effect of Compounds 3a–3h on Cancer Cell Proliferation and Viability of Normal Cells
2.3. Tyrosinase Inhibitory Activity
2.4. Docking Study
3. Materials and Methods
3.1. Chemistry
3.1.1. Synthesis and Structural Characterization
2-(8-Methyl-8-azabicyclo[3.2.1]octan-3-ylidene)hydrazine-1-carbothioamide (2)
4-(4-Iodophenyl)-2-(2-(8-methyl-8-azabicyclo[3.2.1]octan-3-ylidene)hydrazinyl)thiazole (3a)
4-(4-Methoxyphenyl)-2-(2-(8-methyl-8-azabicyclo[3.2.1]octan-3-ylidene)hydrazinyl)thiazole (3b)
4-[2-[2-(8-Methyl-8-azabicyclo[3.2.1]oct-3-ylidene)hydrazinyl]-1,3-thiazol-4-yl]benzonitrile (3c)
4-(4-Nitrophenyl)-2-(2-(8-methyl-8-azabicyclo[3.2.1]octan-3-ylidene)hydrazinyl)thiazole (3d)
4-[2-[2-(8-Methyl-8-azabicyclo[3.2.1]oct-3-ylidene)hydrazinyl]-1,3-thiazol-4-yl]aniline (3e)
N-(4-[2-[2-(8-Methyl-8-azabicyclo[3.2.1]oct-3-ylidene)hydrazinyl]-1,3-thiazol-4-yl]phenyl)methane-sulfonamide (3f)
3-[2-[2-(8-Methyl-8-azabicyclo[3.2.1]oct-3-ylidene)hydrazinyl]-1,3-thiazol-4-yl]-2H-chromen-2-one (3g)
N-(4-[2-[2-(8-Methyl-8-azabicyclo[3.2.1]oct-3-ylidene)hydrazinyl]-1,3-thiazol-4-yl]phenyl)acetamide (3h)
3.2. In Vitro Experiments
3.2.1. Cell Cultures and Drug Treatment
3.2.2. Cell Proliferation Assay
3.2.3. Cell Cytotoxicity Assay
3.2.4. Cell Cycle Progression Assay
3.2.5. Evaluation of Cell Death by Flow Cytometry
3.2.6. Statistical Analysis
3.3. Mushroom Tyrosinase Inhibition Assay
Kinetic Analysis of the Inhibition of Tyrosinase
3.4. Docking Study
3.5. Quantum Calculation
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
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Lipiński’s Rules | Veber’s Rules | Quantum Parameters | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Tropinone | logP | HBA | HBD | MW | NRB | TPSA | EHOMO | ELUMO | ΔE | η | S | IP | EA | χ |
3a | 3.87 | 3 | 1 | 438.33 | 3 | 68.76 | −5.490 | −1.189 | 4.301 | 2.150 | 0.465 | 5.490 | 1.189 | 3.340 |
3b | 3.23 | 4 | 1 | 342.46 | 4 | 77.99 | −5.142 | −0.836 | 4.306 | 2.153 | 0.464 | 5.142 | 0.836 | 2.989 |
3c | 3.30 | 4 | 1 | 337.44 | 3 | 92.55 | −5.734 | −1.791 | 3.943 | 1.971 | 0.507 | 5.734 | 1.791 | 3.762 |
3d | 2.58 | 5 | 1 | 357.43 | 4 | 114.58 | −5.804 | −2.454 | 3.350 | 1.675 | 0.597 | 5.804 | 2.454 | 4.129 |
3e | 2.66 | 3 | 2 | 327.45 | 3 | 94.78 | −4.934 | −0.733 | 4.201 | 2.101 | 0.476 | 4.934 | 0.733 | 2.833 |
3f | 2.41 | 5 | 2 | 405.54 | 5 | 123.31 | −5.463 | −1.188 | 4.274 | 2.137 | 0.468 | 5.463 | 1.188 | 3.326 |
3g | 3.28 | 5 | 1 | 380.46 | 3 | 98.97 | −5.492 | −2.006 | 3.487 | 1.743 | 0.574 | 5.492 | 2.006 | 3.749 |
3h | 2.81 | 4 | 2 | 369.48 | 5 | 97.86 | −5.223 | −0.993 | 4.230 | 2.115 | 0.473 | 5.223 | 0.993 | 3.108 |
Tropinone Derivatives | IC50 ± SD (µM) | CC50 ± SD (µM) | ||||
---|---|---|---|---|---|---|
RPMI 8226 | A549 | MDA-MB-231 | B16-F10 | HSF | CCD-18Co | |
3a | 9.32 ± 0.71 | 13.81 ± 1.90 | 2.26 ± 0.38 | 1.51 ± 0.14 | 19.23 ± 1.77 | 27.76 ± 8.85 |
3b | 42.19 ± 4.55 | 132.7 ± 25.12 | 2.53 ± 0.07 | 2.20 ± 0.04 | 51.46 ± 12.85 | 92.60 ± 1.69 |
3c | 31.70 ± 2.77 | 74.93 ± 7.65 | 2.70 ± 0.14 | 1.52 ± 0.13 | 78.12 ± 1.05 | 101.28 ± 2.61 |
3d | 17.87 ± 1.78 | 30.99 ± 4.62 | 2.44 ± 0.14 | 1.91 ± 0.04 | 35.36 ± 0.97 | 45.57 ± 3.12 |
3e | 67.03 ± 7.14 | 262.9 ± 104.9 | 2.81 ± 0.12 | 2.68 ± 0.40 | nt | nt |
3f | 107.3 ± 7.22 | 117.1 ± 22.82 | 2.48 ± 0.08 | 2.14 ± 0.23 | 98.67 ± 4.16 | nt |
3g | 23.16 ± 4.52 | 29.29 ± 3.98 | 2.83 ± 0.06 | 3.00 ± 0.26 | 35.29 ± 5.52 | 42.58 ± 5.89 |
3h | 107.9 ± 4.80 | 188.8 ± 33.02 | 2.90 ± 0.25 | 3.03 ± 0.18 | nt | nt |
Chlorambucil | - | - | 4.71 ± 0.5 | 2.92 ± 0.10 | - | - |
Compound | IC50 ± SD (µM) | Inhibitory Mechanism | Vmax | KM |
---|---|---|---|---|
3a | 151.44 ± 6.80 | mixed | −325.9 | −515.6 |
3b | 165.38 ± 3.47 | mixed | −280.2 | −409.1 |
3c | 153.04 ± 7.42 | mixed | −753.9 | −914.4 |
3d | 140.21 ± 7.96 | mixed | 4601.9 | 4206.2 |
3e | 118.06 ± 1.77 | mixed | 3392.1 | 3191.3 |
3f | 152.04 ± 4.66 | mixed | −350.2 | −536.4 |
3g | 183.34 ± 6.18 | mixed | −4472.3 | −3295.2 |
3h | 120.71 ± 5.21 | mixed | −271.8 | −433.2 |
Ascorbic acid | 386.5 ± 10.34 | - | - | - |
Compound | Binding Energy ± SD (kcal/mol) |
---|---|
3a | −5.8 ± 0.2 |
3b | −6.4 ± 0.2 |
3c | −7.5 ± 0.3 |
3d | −7.2 ± 0.2 |
3e | −6.5 ± 0.2 |
3f | −6.3 ± 0.1 |
3g | −5.4 ± 0.1 |
3h | −8.1 ± 0.2 |
Ascorbic acid | −4.4 ± 0.6 * |
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Piechowska, K.; Mizerska-Kowalska, M.; Zdzisińska, B.; Cytarska, J.; Baranowska-Łączkowska, A.; Jaroch, K.; Łuczykowski, K.; Płaziński, W.; Bojko, B.; Kruszewski, S.; et al. Tropinone-Derived Alkaloids as Potent Anticancer Agents: Synthesis, Tyrosinase Inhibition, Mechanism of Action, DFT Calculation, and Molecular Docking Studies. Int. J. Mol. Sci. 2020, 21, 9050. https://doi.org/10.3390/ijms21239050
Piechowska K, Mizerska-Kowalska M, Zdzisińska B, Cytarska J, Baranowska-Łączkowska A, Jaroch K, Łuczykowski K, Płaziński W, Bojko B, Kruszewski S, et al. Tropinone-Derived Alkaloids as Potent Anticancer Agents: Synthesis, Tyrosinase Inhibition, Mechanism of Action, DFT Calculation, and Molecular Docking Studies. International Journal of Molecular Sciences. 2020; 21(23):9050. https://doi.org/10.3390/ijms21239050
Chicago/Turabian StylePiechowska, Katarzyna, Magdalena Mizerska-Kowalska, Barbara Zdzisińska, Joanna Cytarska, Angelika Baranowska-Łączkowska, Karol Jaroch, Kamil Łuczykowski, Wojciech Płaziński, Barbara Bojko, Stefan Kruszewski, and et al. 2020. "Tropinone-Derived Alkaloids as Potent Anticancer Agents: Synthesis, Tyrosinase Inhibition, Mechanism of Action, DFT Calculation, and Molecular Docking Studies" International Journal of Molecular Sciences 21, no. 23: 9050. https://doi.org/10.3390/ijms21239050