Exploring N-Ferrocenylmethylaniline Derivatives as Potent Anti-Breast Cancer Agents: Insights from Computational Chemistry, Pharmacokinetics, and QSAR Analysis

1. Exploring N-Ferrocenylmethylaniline Derivatives as Potent Anti-Breast Cancer Agents:
Insights from Computational Chemistry, Pharmacokinetics, and QSAR Analysis
Nadjiba ZEGHEBa,b*, Aicha ADAIKAa, Touhami LANEZa, Tuba Tüylü Küçükkılınç c
a VTRS Laboratory, University of El Oued B.P.789, 39000, El Oued, Algeria.
b Trustlife Labs, Drug Research & Development Center, 34774 Istanbul, Turkey.
cDepartment of Biochemistry, Faculty of Pharmacy, Hacettepe University, Ankara, Turkey
* Corresponding author.Tel.: +213665439433
E-mail address: nadjiba.zegheb@trustlifelabs.com
Introduction
Breast cancer is the most common cancer in women in front
of colorectal cancer and lung cancer. Since the binding of
estradiol to its receptor promotes breast cancer cell proliferation
(in the ER+ tumours), many molecules targeting this protein have
been synthesized to counteract the estradiol action. Ferrocene
derivatives have proved their efficiency against hormone-
dependent breast cancer cells (MCF7). It was found that on MCF-
7 cells, the effects of hydroxyferrocifens are quite similar to that
of hydroxytamoxifen, slightly more potent. Moreover, it was
clarified that the presence of a ferrocene group is necessary to
generate the anti-proliferative effect [1,2].
Objectives of The Study
In this study we aimed to find new N-ferrocenylmethylaniline
(FMA) derivatives having pharmacochemistry properties as
potential drug candidates against human breast cancer cells.
Methodology
Results and Discussion
N°: X Y N°: X Y N°: X Y
A0 H H A10 4-Cl H A20 4-COMe H
A1 H COMe A11 3-Br H A21 H NHCOMe
A2 H COEt A12 4-Br H A22 H NHCOPh
A3 H COPh A13 2-CN H A23 2-NO2 COMe
A4 H CO-Ph-2-NO2 A14 3-CN H A24 3-NO2 COMe
A5 2-Me H A15 4-CN H A25 4-NO2 COMe
A6 3-Me H A16 2-NO2 H A26 2-CN COMe
A7 2-Et H A17 3-NO2 H A27 3-CN COMe
A8 2-Cl H A18 4-NO2 H A28 4-CN COMe
A9 3-Cl H A19 2-COMe H
The synthesis of FMA derivatives (A0-A28) used in this work were
performed following our previously reported procedure[3,4]. Their
molecular structures were fully optimized using DFT/B3LYP with
6-311++(d,p) and LanL2DZ basis sets.
Table 1. General formula of N-ferrocenylmethylaniline derivatives
Results and Discussion
ADME properties
Cytotoxicity MTT assay
Enzyme Result
CYP1A2 A0-A7, A19-A22, A26-A28
CYP2C19 inhibitor All except for A21, A22
CYP2C9 inhibitor All comps. except for A4
CYP2D6 inhibitor All compounds
CYP3A4 inhibitor All comps. except for A4, A16, A23, A24, A25
Figure 3. Docking pose of A27 with ER
illustrating the interactions.
Figure 4. Docking pose of A27 with
PR illustrating the interactions.
Estrogen Receptor Progesterone receptor
Comp
Docking score
(Kcal.mol-1)
Comp
Docking score
(Kcal.mol-1)
A3 -9.85 A3 -10.62
A4 -9.02 A4 -10.13
A22 -9.03 A22 -10.16
A27 -9.05 A24 -10.39
A28 -8.74 A25 -10.15
Table 3. binding free energies (ΔG) with ERα and PR.
2D-QSAR modelling
pIC50= -13.85 +0.0003 E +0.107 MR + 0.022
MV + 0.005 HF - 0.617 Pol -0.029 TPSA -
0.836 qC11 – 0.829 qN1 - 2.179 HOMO
MLR analysis was performed, and the best
model was selected from several QSAR
equations based on different statistical
parameters.
All the values of t-statistics (table 4) are
significant which confirms the significance
of each descriptor. The generated model
was validated for predictive ability inside
the model using Leave One Out method
(LOO) (table5).
R R2 SEE F Q S
0.92 0.84 0.14 9.33 6.75 0.302
Table 4. the statistical parameters of the model
R2
adj R2
cv PE 6PE SSY PRESS SPRESS PRESS/SSY
0.758 0.848 0.021 0.129 2.191 0.332 0.148 0.151
Table 5. Cross-validation parameters
The plot for the model shows a good correlation with experimentally reported data having R² = 0.848. The plot
of residuals against the experimental IC50 values does not show any systematic error.
Graph 2. Scatter Plot between the
Observed and Predicted Activities
of Model.
Graph. 3. Plots of the residual values
against the experimentally observe.
Figure 1. Drug-likeness of
the studied compounds
Figure 2. The boiled egg plot, egg white
represents GI absorption, and the yolk
represents BBB.
Table 2. ADME prediction of the studied compounds
Conclusion & Perspectives
✓The in vitro and in silico studies showed that the studied compounds could be drug candidate against hormone
dependent breast cancer cells.
✓ The obtained QSAR model make it easy to predict the anti-proliferative activity against MCF-7 cell line for
other substituted N-ferrocenylmethylaniline derivatives before carrying any experimental assays.
✓ this work should be further followed by in vitro studies to better investigate their inhibitory effect against
ERs, PR and aromatase.
✓Other MTT assays with prostate cancer cells (PC-3, devoid of ERα but with ERβ) and healthy cell line L929
should be carried out to evaluate the selectivity of compounds on cancer cells over normal cells
References
1. Vessieres, A., et al., Modification of the estrogenic properties of diphenols by the incorporation of ferrocene. Generation of antiproliferative effects in vitro. Journal of medicinal chemistry, 2005.
48(12): p. 3937-3940.
2. Hillard, E., et al., A series of unconjugated ferrocenyl phenols: prospects as anticancer agents. ChemMedChem: Chemistry Enabling Drug Discovery, 2006. 1(5): p. 551-559.
3. Khand, I.U.; Lanez, T.; Pauson P.L. Ferrocene derivatives. Part 24. Synthesis of dihydro-2- pyrindines and dihydro-3H-2- cyclopent[c]azepines by photolysis of thei cyclopentadienyliron
derivatives. J. Chem. Soc.,Perkin Trans. 1, 1989, 2075-2078.
4. Lanez, E.; Bechki, L.; Lanez, T. N6,9- bis(ferrocenylmethyl)adenine: synthesis, cyclic voltammetric, spectroscopic characterization, and DFT calculations, St. Cerc. St. CICBIA, 2019, 20 (4), 509-
519.
5. Zegheb, N., Boubekri, C., Lanez, T., Lanez, E., Küçükkılınç, T. T., Öz, E., ... & Belaidi, S. (2022). In vitro and in silico determination of some N-ferrocenylmethylaniline derivatives as anti-
proliferative agents against MCF-7 human breast cancer cell lines. Anti-Cancer Agents in Medicinal Chemistry (Formerly Current Medicinal Chemistry-Anti-Cancer Agents), 22(7), 1426-1437.
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✓ MCF7
✓ MDA-MB-231
Graph 1. IC50 values of
compounds against
MCF-7 and MDA-MB-
231.
Molecular Docking
Since MDA-MB-231 lacks the estrogen and
progesterone receptors, the molecular
docking was performed on the crystal
structure of estrogen (ERα) and
progesterone (PR) receptors. The overall
results indicated high binding affinity.
Gaussian 09
SwissADME