Pyrazoline B-Paclitaxel or Doxorubicin Combination Drugs Show Synergistic Activity Against Cancer Cells: In silico Study

Figures & data

Figure 1 Visualization of advantages of combination chemotherapy.

Table 1 Adverse Effect of Doxorubicin + Cyclophosphamide Combination. Probability Range: 0–1

Activity	Pa	Pi
DDI arrhythmia	0.000	0.999
DDI bradycardia	0.001	0.985
DDI hypertension	0.000	0.999
DDI hypotension	0.000	0.999
DDI qt_interval_prolongation	0.000	0.999
DDI tachycardia	0.000	0.987

Table 2 Adverse Effect of Doxorubicin + Pyrazoline B Combination. Probability Range: 0–1

Activity	Pa	Pi
DDI arrhythmia	0.018	0.635
DDI bradycardia	0.039	0.504
DDI hypertension	0.002	0.821
DDI hypotension	0.004	0.803
DDI qt_interval_prolongation	0.007	0.761
DDI tachycardia	0.011	0.776

Table 3 Overlapping of P450 Activities of Doxorubicin, Cyclophosphamide, Pyrazoline B. Probability Range: 0–1

	Doxorubicin			Cyclophosphamide			Pyrazoline B
	Inducer	Inhibitor	Substrate	Inducer	Inhibitor	Substrate	Inducer	Inhibitor	Substrate
CYP1A2
CYP2C19						0.574
CYP2C9	0.419		0.916	0.679		0.55
CYP2D6			0.566
CYP3A4	0.507		0.721	0.54	0.677	0.786

Table 4 Adverse Effect of Paclitaxel + Ascorbic Acid Combination. Probability Range: 0–1

Activity	Pa	Pi
DDI arrhythmia	0.005	0.804
DDI bradycardia	0.007	0.836
DDI hypertension	0.001	0.881
DDI hypotension	0.002	0.920
DDI qt_interval_prolongation	0.002	0.934
DDI tachycardia	0.012	0.768

Table 5 Adverse Effect of Paclitaxel + Pyrazoline B Combination. Probability Range: 0–1

Activity	Pa	Pi
DDI arrhythmia	0.036	0.507
DDI bradycardia	0.041	0.491
DDI hypertension	0.004	0.709
DDI hypotension	0.038	0.533
DDI qt_interval_prolongation	0.005	0.787
DDI tachycardia	0.024	0.598

Table 6 Overlapping of P450 Activities of Paclitaxel, Ascorbic Acid, Pyrazoline B. Probability Range: 0–1

	Paclitaxel			Ascorbic Acid			Pyrazoline B
	Inducer	Inhibitor	Substrate	Inducer	Inhibitor	Substrate	Inducer	Inhibitor	Substrate
CYP1A2				0.131		0.152
CYP2C19						0.349
CYP2C9				0.225		0.020
CYP2D6						0.131
CYP3A4			0.793	0.309		0.292

Table 7 Binding Affinity of Pyrazoline B and Doxorubicin Against Target of Doxorubicin (Topoisomerase I and II)

	Binding Affinity (kcal/mol)
	Topoisomerase I	Topoisomerase II
Pyrazoline B	‒7.0	‒7.8
Cyclophosphamide	‒5.1	‒5.7
Doxorubicin	‒8.4	‒9.1

Figure 2 Protein-ligand docking and macromolecular docking of topoisomerase I. (A) The binding of topoisomerase I and pyrazoline B (green) or doxorubicin (red). (B) The binding of topoisomerase I and cyclophosphamide (cyan) or doxorubicin (red). (C) Pyrazoline B-topoisomerase I interaction. (D) Cyclophosphamide-topoisomerase I interaction. (E) Doxorubicin-topoisomerase I interaction. (F) Topoisomerase I without ligand. (G) Topoisomerase I-doxorubicin. (H) Topoisomerase I-pyrazoline B. (I) Topoisomerase I-Doxorubicin-pyrazoline B. (J) Topoisomerase I cyclophosphamide. (K) Topoisomerase I-doxorubicin-cyclophosphamide.

Table 8 Results of Topoisomerase I-DNA Chain Docking

No	Macromolecular Docking	Docking Score	Ligand rmsd (Å)
1	Topoisomerase I-DNA chain	‒509.60	0.48
2	Topoisomerase I doxorubicin-DNA chain	‒499.54	0.34
3	Topoisomerase I pyrazoline B-DNA chain	‒299.96	1.79
4	Topoisomerase I doxorubicin-pyrazoline B-DNA chain	‒301.81	1.87
5	Topoisomerase I cyclophosphamide-DNA chain	‒382.41	0.57
6	Topoisomerase I doxorubicin-cyclophosphamide-DNA chain	‒385.16	0.68

Table 9 Results of Topoisomerase II-DNA Chain Docking

No	Macromolecular Docking	Docking Score	Ligand rmsd (Å)
1	Topoisomerase II-DNA chain	‒936.49	0.70
2	Topoisomerase II doxorubicin-DNA chain	‒937.39	0.72
3	Topoisomerase II pyrazoline B-DNA chain	‒936.43	0.76
4	Topoisomerase II doxorubicin-pyrazoline B-DNA chain	‒936.73	0.70
5	Topoisomerase II cyclophosphamide-DNA chain	‒937.70	0.72
6	Topoisomerase II doxorubicin-cyclophosphamide-DNA chain	‒937.72	0.70

Figure 3 Docking of topoisomerase II and ligands. (A) The binding of topoisomerase II and pyrazoline B (green) or doxorubicin (red). (B) The binding of topoisomerase II and cyclophosphamide (cyan) or doxorubicin (red). (C) Pyrazoline-topoisomerase II interaction. (D) Cyclophosphamide-topoisomerase II interaction. (E) Doxorubicin-topoisomerase II interaction.

Table 10 Binding Affinity of Pyrazoline B and Paclitaxel Against Target of Paclitaxel (α-β Tubulin and Bcl2)

	Binding Affinity (kcal/mol)
	α-β Tubulin	Bcl2
Pyrazoline B	‒6.9	‒7.4
Ascorbic acid	‒6.7	‒4.9
Paclitaxel	‒8.1	‒7.4

Figure 4 Protein-ligand docking and macromolecular docking of α tubulin. (A) Pyrazoline B (green) and Paclitaxel (Orange) binding to α-β tubulin. (B) Ascorbic acid (blue) and Paclitaxel (Orange) binding to α-β tubulin. (C) α-β tubulin-pyrazoline B. (D) α-β tubulin-ascorbic acid. (E) α- β tubulin-paclitaxel. (F) α-tubulin without ligand. (G) α tubulin-Paclitaxel. (H) α tubulin-pyrazoline B. (I) α tubulin-Paclitaxel-pyrazoline B. (J) α tubulin-ascorbic acid. (K) α tubulin-Paclitaxel-ascorbic acid.

Table 11 Macromolecules Docking of α and β Tubulin

No	Macromolecular Docking	Docking Score	Ligand rmsd (Å)
1	α-β tubulin	‒503.57	0.44
2	α tubulin-paclitaxel-β tubulin	‒600.39	0.35
3	α tubulin-pyrazoline B-β tubulin	‒572.05	0.38
4	α tubulin-paclitaxel-pyrazoline B-β tubulin	‒619.53	0.41
5	α tubulin-ascorbic acid-β tubulin	‒564.25	0.38
6	α tubulin-paclitaxel-ascorbic acid –β tubulin	‒612.83	0.45

Figure 5 Protein-ligand docking and macromolecular docking of Bcl2. (A) Pyrazoline B (green) and Paclitaxel (Orange) binding to Bcl2. (B) Ascorbic acid (blue) and Paclitaxel (Orange) binding to Bcl2. (C) Pyrazoline B interaction with residues on Bcl2. (D) Ascorbic acid interaction with residues on Bcl2. (E) Paclitaxel interaction with residues on Bcl2. (F) Bcl2 without ligand. (G) Bcl2-Paclitaxel. (H) Bcl2-pyrazoline B. (I) Bcl2-Paclitaxel-pyrazoline B. (J) Bcl2-ascorbic acid. (K) Bcl2-Paclitaxel-ascorbic acid.

Table 12 Macromolecules Docking of Bcl2 and Bax

No	Macromolecular Docking	Docking Score	Ligand rmsd (Å)
1	Bcl2-Bax	‒510.10	0.30
2	Bcl2-paclitaxel-Bax	‒216.61	13.91
3	Bcl2-pyrazoline B-Bax	‒203.93	12.66
4	Bcl2-paclitaxel-pyrazoline B-Bax	‒204.17	10.93
5	Bcl2-ascorbic acid-Bax	‒454.09	0.41
6	Bcl2-paclitaxel-ascorbic acid-Bax	‒216.72	13.81