Synthesis, Characterization and Biological evaluation of

novel thiosemicarbazide and thiosemicarbazone functionalized

calix[4]arene ligands and related transition metal complexes
Ehsan Bahojb Noruzi, Behrouz Shaabani*, Silvano Geremia, Hossein Samadi Kafil
Faculty of Chemistry, Department of Inorganic Chemistry, University of Tabriz, Tabriz, Iran
Shaabani.b@gmail.com

1. Introduction 3. Results
Calixarenes as the third
generation of supramolecular Ligands L1 and L2 were fully characterized with different
hosts have attracted spectroscopy techniques such as FT-IR, 1H-NMR, 13C-NMR,
considerable attention among
15
N-NMR, COSY, ESI-mass spectroscopy and CHN analysis.
common molecules due to their The complexation of the metal ions was investigated by NMR
easy synthesis from p- titration only for the diamagnetic Zn2+ ion and the preparation
tertbutylphenol and of metal complexes was confirmed by ESI-mass technique.
formaldehyde via base-
catalyzed condensation [1,2]. Co(NO3)2.6H2O
Ni(NO3)2.6H2O
The flexibility of calixarene Cu(NO3)2.3H2O
Zn(NO3)2.4H2O O OH OH O
L1 and L2 + Metal salts O OH OH O and
frame and capability of upper EtOH

rim and lower rim for different HN S S NH

HN S
M
S NH

NH NH
functionalization make them an HN
NH M NH
NH N O O
N

ideal candidate for use as H H

coordinating ligand to different

metal centers. proposed structure for proposed structure for
transition metal complexes of transition metal complexes of Figure 1 : 1H-NMR spectra of ligand L1 and L2
Regarding to complexation (L1) (L2)

capability [3], anti-cancer [4],

antiviral [5] and anti-bacterial
[6] activities we decided to
synthesize a couple of novel
functionalized calix[4]arene
with thiosemicarbazide and
thiosemicarbazone moieties as
ligand and their corresponding
transition metal complexes for
first time.
MTT assays showed considerable in vitro anticancer activity
of Co2+ , Ni2+ and Cu2+ complexes against Saos-2 bone cancer
cell lines. Apoptosis induction properties of the samples upon
treatment of Saos-2 bone cancer cell line were investigated
by microscopic analysis of DAPI stained cells. The untreated
control Saos-2 cells showed normal nuclei and no necrosis of
cancer cells nucleus was observed. Whereas L2 and complex
(Co, Ni and Cu) treated cells showed deformed and fragmented
nuclei in their chromatin and consequently, the morphology of Figure 2: Fluorescent microscopic images of DAPI stained cells following 48
h treatment with A (fresh medium (control)), B (ligand L2), C (complex Co),
the cells was changed. D (complex Ni), E (complex Cu) and F (complex Zn).

2. Experimental
(L1): Calix-NCS (0.15 g, 0.18 mmol) was added at room
temperature to a stirred solution of hydrazine hydrate (90 µL, 1.8
mmol) in ethanol (5 mL). Stirring was continued for 3 h until a
clear yellow solution was obtained. The solvent was evaporated
and the final product was recrystallized from ethanol. Yield 83%.
(L2): A solution of salicylaldehyde (0.48 mL, 44 mmol) in
ethanol (5 mL) was added to a solution of ligand L1 (0.50 g, 5.60
mmol) in ethanol (25 mL). Here after, a yellow powder was
formed. The mixture was refluxed for 6 h and after that the
solution cooled to room temperature, the precipitate was filtered
and washed with cold ethanol to remove unreacted aldehyde.
Yield 78%.
Metal complexes: An appropriate amount of metal salts (0.11
mmol) were dissolved in methanol (5 mL) and the solution was
added to a tetrahydrofuran (THF) solution (10 ml) of ligand (L1
or L2) (0.11 mmol). The mixture was stirred and refluxed for 24
h, after which the precipitate was filtered and the solvent was
eliminated under reduced pressure. The solid obtained was
purified by crystallization using THF.
4. Conclusion
In this study, we designed and synthetized new calix[4]arene based
thiosemicarbazide and thiosemicarbazone molecules and we used them as
four-dentate and six-dentate ligands to prepare a series of transition metal
complexes. The characterization with multiple techniques (FT-IR, 1H-
NMR, 13C-NMR, 15N-NMR, COSY, ESI-Mass spectroscopy, SEM, EDS,
and elemental analysis) demonstrated the success of the synthesis of the
target ligands. To explore the capability of the thiosemicarbazide and
thiosemicarbazone function hosted on a calix[4]arene scaffold for growth
inhibition of bacteria, fungi and cancerous tumor cells (MCF-7, Saos-2 and
MG-63), a series of biological evaluations were performed. Hemolysis
assays demonstrated excellent hemolysis rate for all of the compounds even
at higher concentrations.
5. References
[1] B. Bensenane et. al, Dalton Trans. 45, 2016, 15211-15224.
[2] T. Klejch et. al, New J. Chem. 40, 2016, 7935-7942.
[3] M. Gaber et. al, J. Mol. Struct., 1151, 2018, 56-72.
[4] R. Chen et. al, Molecules, 24, 2019, 2065.
[5] M. Santacruz et. al, Bioorg. & Med. Chemistry, 25, 2017, 4055-4063
[6] M. Molnar et. al, Pharm. Chem. J., 51, 2018, 1078-1081.