Design, Synthesis, and Biological Evaluation of Benzimidazole-Derived Biocompatible Copper(II) and Zinc(II) Complexes as Anticancer Chemotherapeutics
Abstract
:1. Introduction
2. Results and Discussion
2.1. Synthesis and Characterization
2.2. HSA–Metal Complex Interaction by Spectroscopy
2.2.1. Quenching Experiment
2.2.2. Förster Resonance Energy Transfer (FRET) between HSA and Metal Complexes
2.3. Molecular Docking Studies with HSA by Autodock
Prediction of Binding Sites of Metal Complexes on HSA
2.4. DNA Binding/Nuclease Activity
2.5. Molecular Docking Studies with DNA by Hex
Prediction of Binding Sites of Metal Complexes on DNA
2.6. In Vitro Cytotoxicity Assays
2.6.1. Analysis of Growth Inhibition Using 5-Diphenyltetrazolium Bromide (MTT) Assay
2.6.2. Effect on Cancer Cell Adhesion/Migration
2.6.3. Annexin V Apoptosis Detection Assay
2.7. In Vivo Chronic Toxicity Studies of Complex 1 vs. 2
3. Experimental Section
3.1. Material
3.2. Synthesis Procedure of Schiff Base
3.3. Copper Complex: [Cu(BnI)2], (1)
3.4. Zinc Complex, [Zn(BnI)2], (2)
3.5. Fluorescence Quenching Measurements
3.6. FRET Measurements
3.7. DNA Binding and Nuclease Activity
3.8. Molecular Docking Studies
3.8.1. Preparation of Ligands and Receptors
3.8.2. Molecular Docking of Complexes 1 and 2 with DNA
3.8.3. Molecular Docking of Complexes 1 and 2 with HSA
3.9. Cytotoxicity
3.9.1. Cell Lines and Culture Conditions
3.9.2. MTT Assay
3.9.3. Measurement of Cancer Cell Adhesion
3.9.4. Measurement of Cancer Cell Migration
3.9.5. Analysis of Annexin-V Binding by Flow Cytometry
3.10. In Vivo Toxicity
3.10.1. Toxicity Study Design
3.10.2. Chronic Toxicity Study
3.10.3. Hematological Studies
3.10.4. Serum Analysis of Biochemical Parameters
3.10.5. Statistical Analysis
4. Conclusions
Supplementary Materials
Author Contributions
Acknowledgments
Conflicts of Interest
References
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Binding Parameters | |||||
Complex | Temp | KSV | kq | n | Kb |
(K) | ×104 (M−1) | ×1012 (M−1 s−1) | ×104 (M−1) | ||
1 | 298 | 2.02 | 3.54 | 1.09 | 1.58 |
303 | 1.75 | 3.24 | 1.12 | 1.27 | |
308 | 1.51 | 2.64 | 1.20 | 0.86 | |
2 | 298 | 2.74 | 4.79 | 0.99 | 2.78 |
303 | 2.39 | 4.19 | 1.03 | 2.19 | |
308 | 1.97 | 3.45 | 1.11 | 1.48 | |
Thermodynamic Parameters | |||||
Complex | Temp | ∆H | ∆S | T∆S (kcal/mol) | ∆G |
(K) | (kcal/mol) | (cal/mol/K) | (kcal/mol) | ||
1 | 298 | −11.12 | −18.17 | −5.41 | −5.71 |
303 | −5.50 | −5.62 | |||
308 | −5.60 | −5.52 | |||
2 | 298 | −11.48 | −18.14 | −5.40 | −6.08 |
303 | −5.49 | −5.99 | |||
308 | −5.59 | −5.89 | |||
FRET Parameters | |||||
Complex | J | Ro | R | ||
×10−15 (M−1 cm−1) | (nm) | (nm) | |||
1 | 3.26 | 2.04 | 2.35 | ||
2 | 4.63 | 2.16 | 2.36 |
Interaction | Nature of Interaction | Bond Length (Å) | Binding Affinity, Kd (M−1) | ΔG (kcal/mol) |
---|---|---|---|---|
Complex 1 | ||||
Unk:C—Asp451:Oδ2 | Hydrogen Bond | 3.71 | 7.8 × 106 | −9.4 |
Lys199:Cβ—Unk | Hydrophobic (π-σ) | 3.89 | ||
Trp214—Unk | Hydrophobic (π-π) | 4.79 | ||
His242—Unk | Hydrophobic (π-π) | 5.25 | ||
Unk—Trp214 | Hydrophobic (π-π) | 4.91 | ||
Unk—Arg218 | Hydrophobic (π-alkyl) | 5.23 | ||
Unk—Lys195 | Hydrophobic (π-alkyl) | 4.92 | ||
Complex 2 | ||||
Unk:C—His242:Ne2 | Hydrogen Bond | 3.25 | 4.2 × 107 | −10.4 |
Unk:C—Ala291:O | Hydrogen Bond | 3.62 | ||
Lys199:Cβ—Unk | Hydrophobic (π-σ) | 3.68 | ||
Ala291:Cβ—Unk | Hydrophobic (π-σ) | 3.59 | ||
His242—Unk | Hydrophobic (π-π) | 4.86 | ||
His242—Unk | Hydrophobic (π-π) | 4.69 | ||
Unk—Trp214 | Hydrophobic (π-π) | 5.05 | ||
Unk—Lys195 | Hydrophobic (π-alkyl) | 5.02 | ||
Unk—Leu238 | Hydrophobic (π-alkyl) | 5.15 |
Interaction | Bond Distance (Å) | Nature of Interaction | Etotal Value |
---|---|---|---|
Complex 1 | |||
Unk:H—B:d21:O2 | 2.57 | Hydrogen Bond | −238.14 |
Unk:H—B:dG22:O4′ | 2.25 | Hydrogen Bond | |
Unk:H—A:dA5:N3 | 2.34 | Hydrogen Bond | |
Unk:H—A:dA5:N3 | 2.98 | Hydrogen Bond | |
Unk:H—B:d21:O2 | 2.79 | Hydrogen Bond | |
B:d23:OP1—Unk | 4.63 | Electrostatic (π-Anion) | |
Complex 2 | |||
Zn—B:dA18:OP1 | 5.41 | Electrostatic | −248.34 |
Unk:H—A:dG10:O3′ | 2.01 | Hydrogen Bond | |
Unk:H—A:d11:O4′ | 2.89 | Hydrogen Bond | |
Unk:H—A:dG10:OP1 | 2.93 | Hydrogen Bond | |
Unk:H—B:dA18:O3′ | 2.62 | Hydrogen Bond | |
Unk:H—B:dT19:OP1 | 2.81 | Hydrogen Bond | |
B:dA18:OP1—Unk | 2.88 | Electrostatic (π-Anion) | |
B:dA18:OP2—Unk | 4.88 | Electrostatic (π-Anion) | |
B:dA17:O3′—Unk | 2.87 | π-Lone Pair | |
B:dA17:O3′—Unk | 2.55 | π-Lone Pair |
Complex | HepG2 | SK-MEL-1 | HT018 | HeLa | MDA-MB 231 |
---|---|---|---|---|---|
(Liver) | (Skin) | (Colon) | (Cervical) | (Breast) | |
(μM) | (μM) | (μM) | (μM) | (μM) | |
1 | 14 ± 2.2 | 17.8 ± 2.7 | 15 ± 2.1 | 13 ± 2.2 | 3.5 ± 2.4 |
2 | 19 ± 2.6 | 18 ± 2.3 | 25 ± 4.0 | 24.5 ± 2.2 | 26.7 ± 4.1 |
Ligand | NA | NA | NA | NA | NA |
Vehicle control (0.1% DMSO) | NA | NA | NA | NA | NA |
Cisplatin (Positive control) | 6 ± 0.4 | 5.6 ± 0.8 | 5.7 ± 0.2 | 6 ± 0.6 | 3.1 ± 0.2 |
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AlAjmi, M.F.; Hussain, A.; Rehman, M.T.; Khan, A.A.; Shaikh, P.A.; Khan, R.A. Design, Synthesis, and Biological Evaluation of Benzimidazole-Derived Biocompatible Copper(II) and Zinc(II) Complexes as Anticancer Chemotherapeutics. Int. J. Mol. Sci. 2018, 19, 1492. https://doi.org/10.3390/ijms19051492
AlAjmi MF, Hussain A, Rehman MT, Khan AA, Shaikh PA, Khan RA. Design, Synthesis, and Biological Evaluation of Benzimidazole-Derived Biocompatible Copper(II) and Zinc(II) Complexes as Anticancer Chemotherapeutics. International Journal of Molecular Sciences. 2018; 19(5):1492. https://doi.org/10.3390/ijms19051492
Chicago/Turabian StyleAlAjmi, Mohamed F., Afzal Hussain, Md. Tabish Rehman, Azmat Ali Khan, Perwez Alam Shaikh, and Rais Ahmad Khan. 2018. "Design, Synthesis, and Biological Evaluation of Benzimidazole-Derived Biocompatible Copper(II) and Zinc(II) Complexes as Anticancer Chemotherapeutics" International Journal of Molecular Sciences 19, no. 5: 1492. https://doi.org/10.3390/ijms19051492