Synthesis, characterisation, Hirshfeld surface and in vitro cytotoxicity evaluation of new N-aryl-N′-Alkoxycarbonyl thiocarbamide derivatives

Article abstract of DOI:10.1016/j.molstruc.2019.127269

Four new compounds N-(4-nitrophenyl)-N’-(isobutoxycarbonyl) thiocarbamide (1), N-(2, 4-nitrophenyl)-N’-(isobutoxycarbonyl) thiocarbamide (2), N-(4-nitrophenyl)-N’-(ethoxycarbonyl) thiocarbamide (3) and N-(2-Chloro- 4-nitrophenyl)-N’-(ethoxycarbonyl) thiocarbamide (4) were prepared and their structures confirmed by using various spectroscopic (FT-IR, UV–Visible, ¹H and ¹³C NMR) and single crystal X-ray studies of 1 and 3. The presence of intramolecular (N–H?O[dbnd]C) hydrogen bond in the crystal structure of both the compounds causes planarity of carbonyl thiocarbamide unit and trans orientation of C[dbnd]O and C[dbnd]S group. The intermolecular contacts (C–H?S, C–H?O and N–H?S) present in crystal structures have been examined by Hirshfeld surface analysis and their associated 2D fingerprint plots. All the compounds were assessed for their in vitro cytotoxic properties against a panel of seven human cancer cells such as cervical carcinoma (2008, C13*), colorectal (HT29 and HCT116) and ovarian carcinoma (A2780, A2780/CP and IGROV-1). Among them, compounds 2 and 4 exhibited better activity than 1 and 3 against all the cell lines tested.

Full text of DOI:10.1016/j.molstruc.2019.127269

Journal Pre-proof
Synthesis, characterisation, Hirshfeld surface and in vitro cytotoxicity evaluation of
new N-aryl-N′-Alkoxycarbonyl thiocarbamide derivatives
Sunil K. Pandey, Seema Pratap, Sunil K. Rai, Gaetano Marverti, Manpreet Kaur,
Jerry P. Jasinski
PII:
S0022-2860(19)31378-X
DOI:
https://doi.org/10.1016/j.molstruc.2019.127269
MOLSTR 127269
Reference:
To appear in: Journal of Molecular Structure
Received Date: 8 December 2018
Revised Date: 16 October 2019
Accepted Date: 21 October 2019
Please cite this article as: S.K. Pandey, S. Pratap, S.K. Rai, G. Marverti, M. Kaur, J.P. Jasinski,
Synthesis, characterisation, Hirshfeld surface and in vitro cytotoxicity evaluation of new N-aryl-
N′-Alkoxycarbonyl thiocarbamide derivatives, Journal of Molecular Structure (2019), doi: https://
doi.org/10.1016/j.molstruc.2019.127269.
This is a PDF file of an article that has undergone enhancements after acceptance, such as the addition
of a cover page and metadata, and formatting for readability, but it is not yet the definitive version of
record. This version will undergo additional copyediting, typesetting and review before it is published
in its final form, but we are providing this version to give early visibility of the article. Please note that,
during the production process, errors may be discovered which could affect the content, and all legal
disclaimers that apply to the journal pertain.
© 2019 Published by Elsevier B.V.

Graphical Abstract
Synthesis, characterisation, Hirshfeld surface and in vitro cytotoxicity
evaluation of new N-Aryl-N'-Alkoxycarbonyl thiocarbamide derivatives
a
b
c
Sunil K. Pandey , Seema Pratap a*, Sunil K. Rai , Gaetano Marverti , Manpreet Kaur d,
Jerry P. Jasinski d
aDepartment of Chemistry (M.M.V), Banaras Hindu University, Varanasi−221005, UP, India
bOrganic Chemistry Division, CSIR-National Chemical Laboratory (NCL), Pune-411008,
Maharashtra, India
cDepartment of Biomedical, Metabolic and Neural Sciences, University of Modena and Reggio
Emilia, 41125 Modena, Italy
d Department of Chemistry, Keene State College, 229 Main Street, Keene, NH 03435-2001, USA
A series of four N, N'–disubstituted thiocarbamide derivatives (1−4) were synthesized for their
in-vitro anticancer activity evaluation. Significant activity was found in compounds 2 and 4. All
compounds were characterized well through different spectroscopic techniques; however,
structural evaluation of compounds 1 and 3 was done through single crystal X-ray diffraction
data. Further quantitative analysis of noncovalent interactions for compounds 1 and 3 was done

by Hirshfeld surfaces analysis which showed that O⋯H and S⋯H interactions are playing a
major role in stabilizing the crystal lattice.

Synthesis, characterisation, Hirshfeld surface and in vitro cytotoxicity
evaluation of new N-Aryl-N'-Alkoxycarbonyl thiocarbamide derivatives
Sunil K. Pandey a, Seema Pratap a*, Sunil K. Rai b, Gaetano Marverti c, Manpreet Kaur d,
Jerry P. Jasinski d
a
Department of Chemistry (M.M.V), Banaras Hindu University, Varanasi−221005, UP, India
b
Organic Chemistry Division, CSIR-National Chemical Laboratory (NCL), Pune-411008,
Maharashtra, India
c
Department of Biomedical, Metabolic and Neural Sciences, University of Modena and Reggio
Emilia, 41125 Modena, Italy
d
Department of Chemistry, Keene State College, 229 Main Street, Keene, NH 03435-2001,
USA
Abstract
Four new compounds N-(4-nitrophenyl)-N'-(isobutoxycarbonyl) thiocarbamide (1), N-(2, 4-
nitrophenyl)-N'-(isobutoxycarbonyl) thiocarbamide (2), N-(4-nitrophenyl)-N'-(ethoxycarbonyl)
thiocarbamide (3) and N-(2-Chloro- 4-nitrophenyl)-N'-(ethoxycarbonyl) thiocarbamide (4) were
1
prepared and their structures confirmed by using various spectroscopic (FT-IR, UV-Visible, H
and 13C NMR) and single crystal X-ray studies of 1 and 3. The presence of intramolecular
(N−HꢀꢀꢀO═C) hydrogen bond in the crystal structure of both the compounds causes planarity of
carbonyl thiocarbamide unit and trans orientation of C=O and C=S group. The intermolecular
contacts (C−HꢀꢀꢀS, C−HꢀꢀꢀO and N−HꢀꢀꢀS) present in crystal structures have been examined by
Hirshfeld surface analysis and their associated 2D fingerprint plots. All the compounds were
assessed for their in vitro cytotoxic properties against a panel of seven human cancer cells such
as cervical carcinoma (2008, C13*), colorectal (HT29 and HCT116) and ovarian carcinoma
(A2780, A2780/CP and IGROV-1). Among them, compounds 2 and 4 exhibited better activity
than 1 and 3 against all the cell lines tested.
* Email: drseemapratap@gmail.com
Keywords: Thiocarbamide; X-ray crystal structure; In vitro Cytotoxicity; Hirshfeld surface
analysis.

1. Introduction
Since last few decades, cancer has been considered biggest life threatening health problem in the
world. It is a chronic disease characterised by uncontrolled cell proliferation leading to tumor
formation. In order to get good anticancer drug with less side-effects and toxicity a large number
of compounds have been synthesized and tested over the year. N, N'-disubstituted thiocarbamide
derivatives are one such class of compounds exhibiting promising anticancer properties against a
variety of human cancer cell lines [1–5]. The alkoxy thiocarbamide derivatives are considered
one of the popular members in the family which are gaining much attention due to their easy
availability. The remarkable chelating ability of these compounds based on the presence of soft S
and hard N donor atoms in their structural motif facilitate their use as analytical reagents in
liquid–liquid extraction of precious metals (Pt, Pd, Au, Ag) and in extraction of toxic metals
using a solid supported liquid membrane system [6−8]. The technological applications of these
compounds include their use as effective corrosion inhibitors [9], non-linear optical materials in
electronic industries [10], collectors in froth flotation processes [11, 12], precursors for the
synthesis of heterocyclic and organosulfur compounds [1−3], catalyst in oxidation of alcohols
[13, 14] and in the palladium-catalyzed Suzuki and Heck reactions [15−17] and as chemosensors
for selective and sensitive naked-eye recognition for anions [18−20]. The presence of
intramolecular hydrogen bond in the structural scaffold of thiocarbamides has a profound effect
on its conformation and complexation behaviour [13, 14, 21, 22]. In addition, presence of
different types of intermolecular interactions (C−HꢀꢀꢀS, C−HꢀꢀꢀO and N−HꢀꢀꢀS) contributes to the
crystal packing. Hirshfeld surface (HS) analysis can be used to get information about these
interactions [23−26].
Therapeutic importance of these compounds includes their use as herbicidal, fungicidal,
bactericidal, insecticidal, antithyrodal and plant growth regulatory agents [27−31]. They have
been patented as anti-diabetic, anti-arthritic, and anti-coagulant agents for the treatment of
cognitive problem and prostate disorder [32]. Recently substituted thiocarbamide derivatives
have been proved to be most promising anticancer agents because of their good inhibitory
activity against a variety of human cancers [4, 33, 34]. It has also been reported that structural
changes in thiocarbamide moiety greatly affects its anticancer activity [33−35]. In surge to get
better anticancer compounds, our group has synthesized and studied the anticancer properties of
various N, N'-disubstituted thiocarbamides [36−42].

In continuation of our previous work, the present paper describes the “Synthesis,
characterisation, Hirshfeld surface and in vitro cytotoxicity evaluation of four new N-Aryl-N'-
Alkoxycarbonyl thiocarbamide derivatives”.
2. Experimental Procedure
2.1. Chemicals, Instruments and Methods
Analytical grade isobutoxy chloroformate, ethoxy chloroformate and other chemicals used in
synthesis of compounds were purchased from Merck (Germany). The acetone was dried and
freshly distilled prior to use. Melting points were measured on an X–4 digital melting-point
apparatus and were uncorrected. Elemental analyses were performed on a CE-440 Exeter
Analytical CHN analyzer. The infrared spectra of the title compounds as KBr pellets (4000−400
cm-1) were recorded on a Varian 3100 FT-IR Excalibur series spectrophotometer.1H and 13C
NMR spectra were recorded in CDCl3 by using TMS as an internal standard on a JEOL FT-NMR
AL 500 Spectrometer.UV-Visible spectra were recorded on a Shimadzu (-UV-) 1700 Pharma
Spec spectrophotometer, USA. A rectangular quartz cell with optical path length of 1cm was
used.
2.2. X-ray data collection and structure refinement
Data collection was performed using CrysAlisPro on an Oxford Diffraction Xcalibur
diffractometer with a Ruby Gemini CCD detector with graphite - monochromatic MoKα (λ =
0.71073 Å) radiation for compound 1 at 173(2) K and CuKa (λ = 1.54178 Å) radiation for
compound 3 at 293(2) K. The structure was solved by direct methods in the space group and
refined by full-matrix least-square on F2 using SHELXL-97 [43]. The non-hydrogen atoms were
refined with anisotropic thermal parameters. All hydrogen atoms were fixed geometrically and
were allowed refined using a riding model.
2.3. Synthesis of compounds (1−4)
Procedure described in previous literatures was followed for the synthesis compounds after a
slight alteration [3–5]. Isobutoxy chloroformate (1.31 mmol, 30 mL)/ Ethoxy chloroformate
(1.04 mmol, 30 mL) dissolved in dry acetone was mixed to the ammonium thiocyanate solution
(0.78 mmol, 30 mL) in ten minutes duration .The reaction mixture was refluxed for 50 min at 75
°C. The resultant isobutoxy/ethoxy carbonyl isothiocyanate solution was treated with equimolar
quantity of appropriate substituted aromatic primary amine in acetone (Scheme 1) and further

refluxed for 2 h at 30 °C. The by-product ammonium chloride was filtered off and the pale
yellow solutions were evaporated in vacuum to get the desired products. The well-shaped
crystals of compounds 1 and 3 were grown by the slow evaporation techniques using acetone as
a solvent.
Scheme1: Synthetic route to new isobutoxy/ethoxy carbonyl thiocarbamides(1−4).
2. 4. Biological studies
2.4.1. Cell lines
Seven human cancer cell lines namely, cervical (2008 and C13*), colorectal (HT29 and
HCT116) and ovarian carcinoma (IGROV-1, A2780 and A2780/CP) were used. Among these,
C13* and A2780/CP are cisplatin (ccDDP)-resistant cells [44, 45]. Cells were grown as
monolayers in RPMI 1640 medium containing 10% heat-inactivated fetal bovine serum and 50
µg/ml gentamycin sulfate. All cell media and serum were purchased from Lonza (Verviers,
Belgium). Cultures were equilibrated with humidified 5% CO2 in air at 38 ºC. All studies were
performed in Mycoplasma negative cells, as routinely determined with the MycoAlert
Mycoplasma detection kit (Lonza, Walkersville, MD, USA).

2. 4. 2. In vitro cytotoxic screening
In vitro cytotoxicity of compounds was determined by MTT assay [46, 47]. The cells were
seeded into 96-well plates and cultured overnight .Various concentrations of the test compounds
dissolved in DMSO solvents were then added and incubated for 72 h. After incubation, the
medium was removed and added fresh culture medium 100 µl containing 0.5 mg mL-1MTT (3-
(4,5-dimethylthiazol-2-yl)-2,5-diphenytetrazolium bromide; Sigma) and then incubated at 38ºC
for 4 h. The medium was removed and 100 µL DMSO solvent was added to dissolve the dark
blue crystals. After incubation for 30 min at room temperature, to ensure that all crystals were
dissolved, absorbance was measured using an ELISA plate reader at 570 nm with reference
wavelength of 650 nm.
3. Results and discussion
3.1. Synthesis and Characterization
The route adapted for the synthesis of the compounds 1−4 is outline in Scheme 1. The treatment
of isobutoxycarbonyl isothiocyanate/ethoxycarbonyl isothiocyanate with appropriate substituted
aromatic primary amines under reflux conditions afforded light yellow; air stable compounds
1−4 in good yield (79−85%).
3. 2. IR spectra
FT-IR spectral data of all compounds have been presented in SI (sections 2.3.1−2.3.4). The
compounds display two –NH stretching vibrations at ~3440 cm-1(medium) and ~3160 cm-1(
broad) assignable to free ν(NH) and hydrogen bonded ν (NH), respectively. The formation of the
intramolecular N–H⋯O═C hydrogen bond is responsible for the impressive red-shift and strong
intensification of the low frequency NH band [39−42]. This interaction also affects the C═O
1
stretching mode which appears at a lower wave number ( ~1665 cm− ) than that of the ordinary
carbonyl absorption (~1730 cm-1) [41,42]. The medium-intensity absorption observed at ~778
cm-1 is assigned to the ν (C═S) stretching mode. All the assignments are in good agreement with
previously studied N, N'-disubstituted thiocarbamide derivatives [36−42]. The asymmetric and
symmetric −NO2 stretching vibrations of compounds were observed as medium intensity bands
at ~1566 cm-1 and ~1325 cm-1, respectively [41]. In compound 4, C−Cl stretching frequencies
are observed as coupled modes at ~720 cm-1 and ~ 458 cm-1 corresponding to antisymmetric
and symmetric vibrations, respectively [41,42].
3. 3. NMR spectra

13
1H and C spectral data of all compounds have been presented in SI (sections 2.3.1−2.3.4). In
the 1H NMR spectra of the compounds, the protons of the two −NH groups are obtained as single
peak at 12.22−11.76 ppm and 9.10−8.30 ppm, respectively. The presence of intramolecular
hydrogen bonding interaction between CSNH and carbonyl oxygen (N–H⋯O═C) is responsible
for its high chemical shift value [40−42]. All the aliphatic protons and aromatic protons of the
compounds resonate between 1.01−4.19 ppm and 7.52−8.21 ppm, respectively, in their usual
regions. In the 13C NMR spectra of compounds, the chemical shifts of carbons pertaining to
CONH and CSNH moieties resonate at 154.4−148.2 ppm and 178.1−176.0 ppm, respectively.
Signals for aromatic carbons appeared in the region between 140−120 ppm [36−42].
3. 4. UV–Vis Studies
Electronic spectra of compounds 1–4 recorded in dichloromethane over the scan range of 200–
800 nm show identical trends (Figure S1). All compounds present in their absorption spectra a
weak band at around ~260 nm and a strong band at around ~295 nm which can be ascribed to
intraligand charge transfer π → π* transitions [41, 42, 48].
3. 5. Single crystal structure analyses
For compounds 1 and 3 the crystallographic data and refinement information, hydrogen bonds
are gathered in Tables 1 and 2 whereas selected bond lengths, bond angles, torsion angles are
presented in Tables S1 and S2, respectively. The crystal structure, unit cell diagram and packing
pattern of both the compounds are depicted in Figures 1(a, b), S2 (a, b) and 2(a, b), respectively.
In both the compounds the asymmetric unit contains two crystallographically independent but
chemically equal molecules A and B. All the bond lengths and bond angles are almost equal for
the molecules A and B in both the cases. There are only minor differences in the torsion angles
of the thiocarbamide moiety with the aromatic ring (Table S2). C=O and C=S bond lengths
correspond to a double bond whereas all the C−N bonds of thiocarbamide core are the shorter
than the typical single C−N bond length due to delocalization in compounds [49−52]. The
hydrogens at N1 and N2 are anti and syn, respectively, with respect to the C=S bond. Presence of
intramolecular hydrogen bonds C═OꢀꢀꢀH−NC═S and C−HꢀꢀꢀS forming a planar six membered
ring in each case stabilize the crystal structure of the compounds [51−53]. In both the crystals,
the molecules were connected by N−H⋯O, N−H⋯S and C−HꢀꢀꢀO intermolecular hydrogen
bonds with the chain running along b-axis in 1 and 3, respectively, (Figure 2) [51−53].

3. 6. Hirshfeld surface analysis
Hirshfeld surface is the region of a crystal space around the molecule that is defined by a weight
function derived from the atomic electron densities. Therefore, it provides a new way of
exploring packing modes as well as quantitative analysis of the intermolecular interactions in
molecular crystals [54]. Crystal Explorer software is a tool used for the visualizing and exploring
the intermolecular contacts (i.e. hydrogen bonding, vander Waals contacts, C-H⋯π, and π⋯π
stacking) by enabling their 2D fingerprints. These 2D fingerprint plots provide a summary of the
frequency of each combination of de and di across the surface of the molecule and so indicate not
only which interactions are present, but also the relative area of the surface corresponding to
each interaction [54, 55].
The 2D fingerprint plots, bar graph for percentage contributions of various interactions and
Hirshfeld surfaces (dnorm, Shapeindex and Curvedness) are shown in Figures 3 and 4 (decompose
fingerprint plots are shown in Table S3). The asymmetric unit of both the compounds possessed
two independent molecules (i.e. Z’= 2), therefore, Hirshfeld surfaces were generated over both
molecules (i.e. 1 and 3) as well as over individual molecules (i.e. 1A, 1B and 3A, 3B). If we
compare the percentage contribution of polar interactions (N⋯H, O⋯H and S⋯H) in unified
form of both the molecules in the asymmetric unit, compound 1 showed 23.3 % for O⋯H, 13.9
% for S⋯H and 3.4 % for N⋯H while in compound 3 34.4 % for O⋯H, 10.9 % for S⋯H and 1.4
% for N⋯H. Similar order of percentage contribution in individual molecules were also observed
(see the bar graph in Figure 3b). The presence of sharp spikes for O···H and S···H interactions in
2D fingerprints revealed that these interactions are stronger than others, however no such spike
was observed for N···H interaction in any of the molecules. The D···A distance (i.e. di + de) for
O···H interactions contributed by C‒H···O interactions were ~2.45 Å for compound 1, however,
it was ~2.30 Å for compound 3. This showed that C‒H···O interactions are stronger in
compound 3 than 1. The D···A distances (i.e. di + de) for S···H interactions contributed by N‒
H···S and C‒H···S, both in compound 1 were ~2.40 Å, however, it was ~2.30 Å for compound 3
and it was contributed by only N‒H···S interactions. The percentage contribution due to
aromatic interactions (i.e. C‒H···π and π···π) were insignificant. Although H···H interactions
were 10.9 % and 13.9 % in compounds 1 and 3, respectively. The absence of side wings in the
fingerprints indicated that there was no contribution of C‒H···π interactions in C···H contacts.

However, the presence of bright bins in the fingerprint of compound 3 indicated the contribution
of π···π interactions in C···C contacts [23−26].
In compound 1 dnorm showed six short contacts (i.e. red colour) over each molecule whereas 3
showed five short contacts (Figure 3a). These prominent short contacts were belonging to the
S···H and O···H interactions. But we cannot identify that underlying species is donor or
acceptor. The nature of contacts (i.e. either donor or acceptor) on the Hirshfeld surfaces could
easily be identified by mapping on a shape index. The area over the acceptor was red and donor
was blue over shape index. Similarly intermolecular stacking can easily be identified by
curvedness. The curvedness plots showed that flat surface patches were more prominent in
molecule 1 than 3 (Figure 4a, b). It indicates that diversion of intermolecular planes in crystal
lattice of compound 1 was modest however in 3 it was significant [25, 26].
3. 7. In vitro cytotoxicity
The anticancer activity of the synthesized compounds were assayed against a panel of seven
human cancer cell lines, including cervical (2008 and C13*), colorectal (HT29 and HCT116) and
ovarian carcinoma (IGROV-1, A2780 and A2780/CP) and the results expressed as IC50 are
summarized in Table 3. 5-Flourouracil was used as control [56−58]. The comparative
cytotoxicity of the compounds is depicted in Figure 5. In general, the compounds exhibited good
inhibitory properties for all the cell lines tested. It can be inferred from Table 3 that all the
compounds showed comparatively better activity against cervical (2008 and C13*) and ovarian
carcinoma (IGROV-1) cell lines in accordance with our previous results [36−42]. In comparison
to 1 and 3, Compound 2 and 4 exhibited strong inhibitory activity against all the cell lines which
might be due to the presence of two nitro and/or chloro electron withdrawing group at ortho and
para positions [59, 60]. Previous studies have shown that the presence of electronegative
atom/group at ortho and or/ para position of the aromatic ring increases the lipophilicity of
molecules and is responsible for enhanced cytotoxicity in MTT model [4, 5]. In general, there is
not much change in the inhibitory activity of these compounds in comparison to our previously
reported ones [36−42]. Based on the above results, potential anticancer candidates can be
considered for structural modification and pharmacological evaluation.
4. Conclusions
In the present work, four novel N, N'-disubstituted thiocarbamides have been synthesized and
characterized using various spectroscopic and single crystal X-ray studies. From our experimental

results, it can be concluded that compounds adopt syn-anti planar conformation by virtue of the
presence of strong intramolecular C=O···H–N hydrogen bond interaction in both solution and
solid state. Also, we report detailed analysis of intermolecular interactions (C−HꢀꢀꢀO and
N−HꢀꢀꢀS) present in compounds by Hirshfeld surfaces analysis and associated fingerprint plots of
compounds 1 and 3. In vitro cytotoxicity results of the synthesized compounds against a panel of
seven human cancer cell lines revealed them to be potential inhibitors of cervical (2008, C13*)
and ovarian (IGROV-1) cancer cells. In addition, the better inhibitory activity of compounds 2
and 4 than 1 and 3 may be credited to the presence of two electron-withdrawing groups (Cl and
NO2) on the phenyl ring.
Acknowledgements
The author, SKP is grateful to University Grant Commission, New Delhi, India for the financial
assistance. I am thankful to Prof. G. Marverti, Department of Biomedical Sciences, Metabolic
and neurosciences, University of Modena, 41125 Modena, Italy for cytotoxicity studies. I am
also very thankful to Prof. Jerry P. Jasinski, Department of Chemistry, Keene State College, and
229 Main Street, Keene, NH 03435-2001, USA for single crystal X-ray diffraction. SKR thanks
SERB, New Delhi, for National Postdoctoral Fellowship.
Supporting Information Summary
Detailed experimental procedure and spectral data of all compounds (Sections 2.3.1−2.3.4).
UV−Vis spectra of compounds (1−4) in Figure S1, Unit cell diagram of compounds 1 and 3 have
been provided in Figure S2 and FT-IR spectra of compounds (1−4) in Figure S3. Selected bond
lengths (Å) and bond angles (°) for compounds 1 and 3 in Table S1, Selected torsion angles (°)
for compounds 1 and 3 in Table S2 and Various weak interactions in compound 1 and 3 in
Table S3. Scan copies of 1HNMR Figures (S4−S7) and 13CNMR Figures (S8−S11).
CCDC 1411613 and 1570186 contain the supplementary crystallographic data for compounds
(1) and (3). These data can be obtained free of charge from The Cambridge Crystallographic
Data Centre via www.ccdc.cam.ac.uk/data_request/cif, by e-mailing deposit@ccdc.cam.ac.uk or
by contacting the Cambridge Crystallographic Data.

References:
[1] A. Saeed, U. Flörke, M. F. Erben, A review on the chemistry, coordination, structure and
biological properties of 1-(acyl/aroyl)-3-(substituted) thioureas, J. Sulfur Chem. 35 (2014)
318−355.
[2] A. A. Aly, E. K. Ahmed, K. M. El-Mokadem, M. El-Amir, F. Hegazy, Update survey on
aroyl substituted thioureas and their applications, J. Sulfur Chem. 28 (2007) 73−93.
[3] K. R. Koch, New chemistry with old ligands: N-alkyl- and N, N-dialkyl-N'-acyl (aroyl)
thioureas in co-ordination, analytical and process chemistry of the platinum group metals, Coord.
Chem. Rev. 216–217 (2001) 473−488.
[4] S. Saeed, N. Rashid, P. G. Jones, M. Ali, R. Hussain, Synthesis, characterization and
biological evaluation of some thiourea derivatives bearing benzothiazole moiety as potential
antimicrobial and anticancer agents, Eur. J. Med. Chem. 45 (2010) 1323−1331.
[5] O. E. Hernández, J. Duque, E. Reguera, Structural features of 1-furoylthioureas 3-
monosubstituted and 3, 3-disubstituted: coordination to cadmium and analytical applications, J.
Sulfur Chem. 32 (2011) 213−222.
[6] M. Ilişa, M. Micutzb, I. Pasukc, T. Staicub, V. Cîrcua, Synthesis and liquid crystalline
properties of novel fluorinated N-benzoyl thiourea compounds. Effect of perfluoroalkyl chains
on the thermal behavior and smectic phases stability, J. Fluorine Chem. 204 (2017) 84–89.
[7] T. Seshadri, H. J. Haupt, Thermotropic properties of monosubstituted ferrocene derivatives
bearing bidentate N-benzoyl-N'-arylthiourea ligands-novel building blocks for heterometallic
liquid crystal systems, J. Mater. Chem. 8(6) (1998) 1345–1350.
[8] M. M. Habtu, S. A. Bourne, K. R. Koch, R. C. Luckay, Competitive bulk liquid membrane
transport and solvent extraction of some transition and post-transition metal ions using
acylthiourea ligands as ionophores, New J. Chem. 30 (2006) 1155–1162.
[9] C. G. M. de Oliveira, V. W. Faria, G. F. de Andrade, E. D. Elia, M. F. Cabral, B. A. Cotrim,
G. O. Resende, F. C. de Souza, Synthesis of Thiourea Derivatives and its Evaluation as
Corrosion Inhibitor For Carbon Steel, Phosphorus, Sulfur, and Silicon 190 (2015) 1366–1377.
[10] V. Krishnakumar, C. Ramachandraraja, R.S. Sundararajan, Crystal growth and vibrational
spectroscopic studies of the semiorganic non-linear optical crystal-bisthiourea magnesium
sulphate, Spectrochim. Acta Part A 68 (2007) 113–116.

[11] L. G. yi, Z. Hong, X. Liu-yin, W. Shuai, X. Zheng-he, Improving copper flotation recovery
from a refractory copper porphyry ore by using ethoxycarbonyl thiourea as a collector, Min. Eng.
24 (2011) 817–824.
[12] G.A. Hope, R. Woods, S.E. Boyd, K. Watling, A SERS spectroelectrochemical
investigation of the interaction of butylethoxycarbonylthiourea with copper surfaces, Collo.
Surfaces A: Physicochem. Eng. Aspects 232 (2004) 129–137.
[13] N. Gunasekaran, P. Ramesh, M. N. G. Ponnuswamy, R. Karvembu, Monodentate
coordination of N-[di(phenyl/ethyl)carbamothioyl] benzamide ligands: synthesis, crystal
structure and catalytic oxidation property of Cu(I) complexes, Dalton Trans. 40 (2011)
12519−12526.
[14] M. M. Sheeba, M. M. Tamizh, L. J. Farrugia, A. Endo, R. Karvembu, Chiral
(η6 p Cymene)ruthenium(II) Complexes Containing Monodentate Acylthiourea Ligands for
Efficient Asymmetric Transfer Hydrogenation of Ketones, Organometallics.33 (2014) 540−550.
[15] D. Yang, Y. C. Chen, N. Y. Zhu, Sterically Bulky Thioureas as Air - and Moisture-Stable
Ligands for Pd-Catalyzed Heck Reactions of Aryl Halides, Org. Lett. 6 (2004) 1577−1580.
[16] T. Y. Zhang, M. J. Allen, An Easily Prepared, Air and Moisture Stable, Resin-bound
Palladium Catalyst for Suzuki Cross-Coupling Reactions, Tetrahedron Lett. 40 (1999)
5813−5816.
[17] D. P. Singh, S. Pratap, S. K. Gupta, R. J. Butcher, Structural and spectral speciation on
methyl 2-(3-(furan-2-carbonyl) thioureido) benzoate: A comparative experimental and
theoretical study, J. Mol. Struct. 1048 (2013) 500–509.
[18] S. Li, X. Cao, C. Chen, S. Ke, Novel salicylic acid-oriented thiourea-type receptors as
colorimetric chemosensor: Synthesis, characterizations and selective naked-eye recognition
properties, Spectrochim. Acta Part A: 96 (2012) 18–23.
[19] S. Hua, Y. Guo, J. Xu, S. Shao, Tunability of anion binding strength based on acyl-thiourea
receptors containing isatin group, Spectrochim. Acta Part A: 72 (2009) 1043–1046.
[20] M. Bonizzoni, L. Fabbrizzi, A. Taglietti, F. Tiengo, (Benzylideneamino) thioureas –
Chromogenic Interactions with Anions and N–H Deprotonation, Eur. J. Org. Chem. (2006)
3567–3574.

[21] S. Saeed, R. Hussain, Preparation and characterization of copper sulfide nanoparticles from
symmetrical [(Bu)2NC(S)NC(O)C6H3(3,5-NO2)2]2Cu(II) and [(Bu)2NC(S)NC(O)C6H4(4-
NO2)]2Cu(II) complexes by thermolysis, J. Coord. Chem. 67 (2014) 2942−2953.
[22] S. S. Hassan, M. M. Shoukry, R. N. Shallan, R. van Eldik, Synthesis, characterization,
speciation and biological studies on metal chelates of 1- benzoyl(1,2,4-triazol-3-yl)thiourea, J.
Coord. Chem.70 (2017) 1761−1775.
[23] A. Saeed, M. Bolte, M. F. Erben, H. Pérez, Intermolecular interactions in crystalline 1-
(adamantane-1-carbonyl)-3-substituted
CrystEngComm,17 (2015) 7551–7563.
thioureas
with
Hirshfeld
surface
analysis,
[24] L. Qiao, Y. Zhang, W. Hu, J. Guo, W. Cao, Z. Ding, Z. Guo, A. Fan, J. Song, J. Huang,
Synthesis, structural characterization and quantum chemical calculations on 1-(isomeric methyl
benzoyl)-3-(4-trifluoromethylphenyl) thioureas, J. Mol. Struct. 1141 (2017) 309-321.
[25] L. Qiao, J. Huang, W. Hu, Y. Zhang, J. Guo, W. Cao, K. Miao, B. Qin, J. Song, Synthesis,
characterization, and in vitro evaluation and in silico molecular docking of thiourea derivatives
incorporating 4-(trifluoromethyl) phenyl moiety, J. Mol. Struct. 1139 (2017) 149-159.
[26] I. Gumus, U. Solmaz, G. Binzet, E. Keskin, B. Arslan, H. Arslan, Hirshfeld surface analyses
and crystal structures of supramolecular self-assembly thiourea derivatives directed by non-
covalent interactions,J. Mol. Struct. 1157 (2018) 78-88.
[27] S. Tahir, A. Badshah, R. A. Hussain, M. N. Tahir, S. Tabassum, J. A. Patujo, M. K. Rauf,
DNA-binding studies and biological activities of new nitrosubstituted acyl thioureas, J. Mol.
Struct. 1099 (2015) 215−225.
[28] A. Sethukumar, C. U. Kumar, R. Agilandeshwari, B. A. Prakasam, Synthesis,
stereochemical, structural and biological studies of some 2,6-diarylpiperidin-4-one N(4')-
cyclohexyl thiosemicarbazones, J. Mol. Struct. 1047 (2013) 237–248.
[29] A. Saeed, S. Zaib, S. Ashraf, J. Iftikhar, M. Muddassar, Kam Y.J. Zhang, J. Iqbal, Synthesis,
cholinesterase inhibition and molecular modelling studies of coumarin linked thiourea
derivatives, Bioorg. Chem. 63 (2015) 58–63.
[30] J. N. Asegbeloyin, E. E. Oyeka, O. Okpareke, A. Ibezim, Synthesis, structure,
computational and in-silico anticancer studies of N, N-diethyl-N'-palmitoyl thiourea, J. Mol.
Struct. 1153 (2018) 69−77.

[31] A. P. Keche, G. D. Hatnapure, R. H. Tale, A. H. Rodge, V. M. Kamble, Synthesis, anti-
inflammatory and antimicrobial evaluation of novel 1-acetyl-3,5-diaryl-4,5-dihydro (1H)
pyrazole derivatives bearing urea, thiourea and sulfonamide moieties, Bioorg. Med. Chem. Lett.
22 (2012) 6611–6615.
[32] C. J. E. Haynes, N. Busschaert, I. L. Kirby, J. Herniman, M. E. Light, N. J. Wells, I.
Marques, V. Félix, P. A. Gale, Acylthioureas as anion transporters: the effect of intramolecular
hydrogen bonding, Org. Biomol. Chem. 12 (2014) 62–72.
[33] G. Hallur, A. Jimeno, S. Dalrymple, T. Zhu, M. K. Jung, M. Hidalgo, J. T. Isaacs, S.
Sukumar, E. Hamel, S. R. Khan, Benzoylphenylurea Sulfur Analogues with Potent Antitumor
Activity, J. Med. Chem. 49 (2006) 2357−2360.
[34] S.N. Manjula , N. M. Noolvi, K. V. Parihar, S.A. M. Reddy, V. Ramani, A. K. Gadad, G.
Singh, N. G. Kutty, C. M. Rao, Synthesis and antitumor activity of optically active thiourea and
their 2-aminobenzothiazole derivatives: A novel class of anticancer agents, Eur. J. Med. Chem.
44 (2009) 2923−2929.
[35] F. A. Larik, M. Faisal, A. Saeed, P. A. Channar, J. Korabecny, F. Jabeen, I. A. Mahar, M.
A. Kazi, Q. Abbas, G. Murtaza, G. S. Khan, M. Hassan, S.Y. Seo, Investigation on the effect of
alkyl chain linked mono-thioureas as Jack bean urease inhibitors, SAR, pharmacokinetics
ADMET parameters and molecular docking studies, Bio. Chem. 86 (2019) 473–481.
[36] D. P. Singh, S. Pratap, S. K. Pandey, R. J. Butcher, G. Marverti, N-(naphthyl)-N′-
(methoxy carbonyl) thiocarbamide and its Cu (I) complex: synthesis, spectroscopic, X-ray, DFT
and in vitro cytotoxicity study, J. Coord. Chem. 68 (2015) 261−276.
[37] D. P. Singh, S. Pratap, M. Shukla, Solvent induced geometry transformation of trigonal
planar Cu(I) complexes of N-((2/4-methyoxy carbonyl) phenyl)-N'-(ethoxy/methoxy carbonyl)
thiocarbamides to square-planar Cu(II) complexes: Synthesis, spectral, single crystal, DFT and in
vitro cytotoxic study, Inorg. Chim.Acta. 423 (2014) 386−396.
[38] S. K. Pandey, D. P. Singh, G. Marverti, R. J. Butcher, S. Pratap, Monodentate Coordination
of N, N’-Disubstituted Thiocarbamide Ligands: Syntheses, Structural Analyses, In Vitro
Cytotoxicity and DNA Damage Studies of Cu(I) Complexes, ChemistrySelect 3 (2018)
3675−3679.

[39] S. K. Pandey, S. Pratap, G. Gozzi, G. Marverti, R. J. Butcher, Synthesis, molecular structure
exploration and in vitro cytotoxicity screening of five novel N, N′-disubstituted thiocarbamide
derivatives, Phosphorus, Sulfur, and Silicon Relat. Elem. 193 (2018) 507−514.
[40] S. K. Pandey, S. Pratap, M. K. Tiwari, G. Marverti, J. P. Jasinski, Experimental and
theoretical exploration of molecular structure and anticancer properties of two N, N'-
disubstituted thiocarbamide derivatives, J. Mol. Struct. 1175 (2019) 963−970.
[41] S. K. Pandey, S. Pratap, G. Marverti, M. Kaur, J. P. Jasinski, Synthesis, spectroscopic,
crystal structure and in vitro cytotoxicity studies of N thiophenoyl- N′-substituted phenyl
thiocarbamide derivatives, J. Mol. Struct. 1180 (2019) 447−454.
[42] S. K. Pandey, S. Pratap, S.K. Rai, G. Marverti, M. Kaur, J. P. Jasinski, Synthesis,
characterization, Hirshfeld surface, cytotoxicity, DNA damage and cell cycle arrest studies of N,
N-diphenyl-N'-(biphenyl-4-carbonyl/4-chlorobenzoyl) thiocarbamides J. Mol. Struct.1186 (2019)
333−344.
[43] L.J. Farrugia, WinGX and ORTEP for windows: an update, J. Appl.Cryst. 45 (2012) 849-
854.
[44] C. Korch, M. A. Spillman, T.A. Jackson, B. M. Jacobsen, S. K. Murphy, B. A. Lessey, V.
C. Jordan, A. P. Bradford, DNA profiling analysis of endometrial and ovarian cell lines reveals
misidentification, redundancy and contamination, Gynecol Oncol. 127 (2012) 241−248.
[45] A. P. Andrews, A. J. Jones, Characterization of Binding Proteins from Ovarian Carcinoma
and Kidney Tubule Cells that are Specific for Cisplatin Modified DNA, Cancer Commun. 3
(1991) 93−102.
[46] T. Mosmann, Rapid colorimetric assay for cellular growth and survival: Application to
proliferation and cytotoxicity assays, J. Immunol. Methods. 65 (1983) 55−63.
[47] A. A. Stepanenko, V.V. Dmitrenko, Pitfalls of the MTT assay: Direct and off-target effects
of inhibitors can result in over/underestimation of cell viability, Gene 574 (2015) 193−203.
[48] A. Saeed, S. Ashraf, J. M. White, D. B. Soria, C. A. Franca, M. F. Erben, Synthesis, X-ray
crystal structure, thermal behavior and spectroscopic analysis of 1-(1-naphthoyl)-3-(halo-
phenyl)-thioureas complemented with quantum chemical calculations, Spectrochim. Acta Part A.
150 (2015) 409–418.

[49] F.H. Allen, O. Kennard, D.G. Watson, L. Brammer, A.G. Orpen, R. Taylor, Tables of bond
lengths determined by X-ray and neutron diffraction. Part 1. Bond lengths in organic compounds,
J. Chem. Soc. Perkin Trans. 2 (12) (1987) S1-S19.
[50] A. Saeed, M. S. Shah, F. A. Larik, S. U. Khan, P. A. Channar, U. Flörke, J. Iqbal,
Synthesis, computational studies and biological evaluation of new 1-acetyl-3-aryl thiourea
derivatives as potent cholinesterase inhibitors, Med. Chem. Res. 26 (2017) 1635–1646.
[51] R. I. Al-Wabli, A. Salman, V. Shyni, H. A. Ghabbour, I. H. Joe, M. S. Almutairi, Y. A.
Maklad, M. I. Attia, Synthesis, crystal structure, vibrational profiling, DFT studies and molecular
docking of N-(4-chloro-2-{[2-(1H-indol-2-ylcarbonyl) hydrazinyl] (oxo) acetyl}phenyl)
acetamide. DMSO: A new antiproliferative agent, J. Mol. Struct. 1155 (2018) 457-468.
[52] F. A. Larik, M. S. Shah, A. Saeed, H. S. Shah, P. A. Channar, M. Bolte, J. Iqbal, New
cholinesterase inhibitors for Alzheimer's disease: Structure activity relationship, kinetics and
molecular docking studies of 1–butanoyl–3–arylthiourea derivatives, Int. J. Bio. Macromol. 116
(2018) 144–150.
[53] E. C. Aguilar, G. A. Echeverría, O. E. Piro, S. E. Ulic, J. L. Jios, M. E. Tuttolomondo, H.
Pérez, Weak and strong hydrogen bonds conducting the supramolecular framework of 1-butyl-3-
(1-naphthoyl) thiourea: crystal structure, vibrational studies, DFT methods, Pixel energies and
Hirshfeld surface analysis, Mol. Physics, 116 (2018) 399−413.
[54] J. J.McKinnon, M. A. Spackmana, A. S. Mitchell, Novel tools for visualizing and exploring
intermolecular interactions in molecular crystals, Acta Cryst. B60 (2004) 627-668.
[55] J. J. McKinnon, D. Jayatilaka, M. A. Spackman, Towards quantitative analysis of
intermolecular interactions with Hirshfeld surfaces, Chem. Commun. (2007) 3814–3816.
[56] A. A. Ramos, C. P. Wilsonb, A. R. Collins, Protective effects of Ursolic acid and Luteolin
against oxidative DNA damage include enhancement of DNA repair in Caco-2 cells, Mutat. Res.
692 (2010) 6−11.
[57] K. K. Gnanasekaran, D. M. Benbrook, B. Nammalwar, E. Thavathiru, R. A. Bunce, K. D.
Berlin, Synthesis and evaluation of second generation Flex-Het scaffolds against the human
ovarian cancer A2780 cell line, Eur. J. Med. Chem. 96 (2015) 209−217.
[58] K. M. Khan, F. Naz, M. Taha, A. Khan, S. Perveen, M. I. Choudhary, W. Voelter, Synthesis
and in vitro urease inhibitory activity of N,N'-disubstituted Thioureas, Eur. J. Med. Chem. 74
(2014) 314−323.

[59] H. N. Pati, U. Das, J. W. Quail, M. Kawase, H. Sakagami, J. R. Dimmock, Cytotoxic 3,5-bis
(benzylidene) piperidin-4-ones and N-acyl analogs displaying selective toxicity for malignant
cells, Eur. J. Med. Chem. 43 (2008) 1−7.
[60] M. K. Rauf, I. Din, A. Badshah, M. Gielen, M. Ebihara, D. deVos, S. Ahmed, Synthesis,
structural characterization and in vitro cytotoxicity and anti-bacterial activity of some copper(I)
complexes with N, N'-disubstituted thioureas, J. Inorg. Biochem. 103 (2009) 1135–1144.

Table 1: Crystal data and structure refinement for compounds 1 and 3.
Compounds
1
3
CCDC No.
Empirical Formula
Formula Weight
1411613
C12H15N3O4S
297.33
1570186
C10H11N3O4S
269.28
Crystal System / Space Group
a / Å
b / Å
c / Å
α / °
Triclinic / P-1
10.6343(6)
11.2933(6)
12.5340(6)
89.576(4)
75.582(4)
73.192(5)
1392.21(13)
4
Triclinic / P-1
8.0975(5)
11.7854(8)
13.9092(9)
73.746(6)
74.419(6)
84.785(5)
1227.32(15)
4
β / °
γ / °
V / Å3
Z
D calc (g/cm3)
1.419
1.457
ꢀ (mm-1)
F(000)
0.249
624.0
0.39 × 0.36 × 0.34
MoKα (λ = 0.71073)
2.481
560.0
0.22 × 0.06 × 0.04
CuKα (λ = 1.54184)
Crystal size (mm3)
Radiation
Color / Shape
Temp (K)
Theta range for collection
Index ranges
Bright yellow single crystal Bright yellow single crystal
173(2)
5.972 to 65.64
293(2)
6.844 to 142.586
-7 ≤ h ≤ 9, -14 ≤ k ≤ 13, -
14 ≤ l ≤ 16
-16 ≤ h ≤ 8, -17 ≤ k ≤ 16, -
19 ≤ l ≤ 18
Reflections collected
16672
8213
Independent reflections
9146 [Rint = 0.0308,
Rsigma = 0.0476]
9146/0/365
4626 [Rint = 0.0316,
Rsigma = 0.0427]
4626/0/327
Data/restraints/parameters
Goodness of fit on F2
1.014
1.031
Final R indices [I > 2σ(I)]
R indices (all data)
Largest difference peak/hole
R1 = 0.0447, wR2 = 0.1049
R1 = 0.0681, wR2 = 0.1212
0.34/-0.26 Å-3
R1 = 0.0478, wR2 = 0.1259
R1 = 0.0698, wR2 = 0.1418
0.27/-0.24 Å-3

Table 2: Selected weak hydrogen bonding intermolecular interactions for compounds 1 and 3
(Å°).
D−H···A
Compound 1
D−H
H···A
D···A
<(D−H−A)
N1B−H1BꢁꢁꢁS1A #1
N2B−H2BꢁꢁꢁO2B
0.88
0.88
0.95
0.88
0.88
0.95
0.95
2.53
1.89
2.55
2.62
1.93
2.57
2.51
3.3677(11)
2.6457(15)
3.2261(14)
3.4502(11)
2.6688(15)
3.4162(16)
3.1930(14)
159.3
142.9
128.5
157.6
140.7
148.9
128.5
C12B−H12BꢁꢁꢁS1B
N1A−H1AꢁꢁꢁS1B #1
N2A−H2AꢁꢁꢁO2A
C9A−H9AꢁꢁꢁO2A#2
C12A−H12AꢁꢁꢁS1A
Compound 3
N1A−H1AꢁꢁꢁO1A
N2A−H2AꢁꢁꢁS1B #1
C3A−H3ABꢁꢁꢁO3B#2
C9A−H9AꢁꢁꢁO1B#2
N1B−H1BꢁꢁꢁO1B
N2B−H2BꢁꢁꢁS1A#1
C3B−H3BBꢁꢁꢁO3B #2
C6B−H6BꢁꢁꢁS1B
0.86
0.86
0.97
0.93
0.86
0.86
0.97
0.93
0.93
0.93
1.96
2.47
2.61
2.59
1.95
2.56
2.59
2.47
2.63
2.47
2.673(3)
3.299(2)
3.216(3)
3.396(3)
2.678(3)
3.401(19)
3.479(4)
3.147(2)
3.401(3)
3.366(3)
140.1
161.5
121.0
146.0
141.3
167.7
151.8
129.9
141.0
161.0
C7B−H7BꢁꢁꢁO4A #3
C9B−H9BꢁꢁꢁO1A #2
Symmetry transformations used to generate equivalent atoms:
1. #1. -X, 1-Y, -Z. #2. –X, -Y, 1-Z
3. #1.1-X, 2- Y, 1-Z; #2. 2-X, 1-Y, 1-Z. #3. 1-X, 1-Y, 2-Z

Table 3: IC50 values (µM) for the compounds 1−4 against seven human cancer cell lines;
(cervical 2008 and C13*), (colorectal HT29 and HCT116) and (ovarian carcinoma A2780,
A2780/CP and IGROV-1)
Comp 2008
ounds
C13*
HT29
HCT116
A2780
A2780/CP
IGROV-1
38.0±0.2 31.2±0.1 44.6±1
45.6±0.5
49.6±0.4
34.6±0.5
39.1±0.3
33.1±0.8
5.5±0.3
42.5±0.3
38.1±1
32.2±0.2
28.6±5
1
27.7±0.3 25.3±0.9 37.3±0.9 39.1±0.1
2
28.7±1
22.1±0.5 23.4±0.7 31.6±2
4.1±0.3
24.4±2
30.6±4
33.7±0.5
29.1±0.8
36.2±0.6
35.5±1
27.6±0.3
21.6±0.7
5.1±0.2
3
4
8.5±0.5 15.2±2.1 13.5±1.8
12.8±0.5
5-FU

(a)
(b)
Fig. 1 ORTEP views of (a) Compound 1 and (b) Compound 3 with probability thermal ellipsoids
at the 50% level.

(a)

(b)
Fig. 2 Crystal packing of (a) Compound 1 and (b) Compound 3 viewed along the b-axis.

Fig. 3 (a) 2D fingerprint plot of compounds 1 and 3, (1A, 1B and 3A, 3B represent each
individual molecules in asymmetric unit of 1 and 3, respectively) (b) Percentage contribution of
various intermolecular interactions obtained from decomposed fingerprint plots.
Fig. 4 (a) and (b) represent the Hirshfeld surfaces mapped over dnorm, Shape Index and
Curvedness for compounds
1 and 3, respectively.

Fig. 5 Comparative in vitro cytotoxicity of compounds 1−4 for a panel of seven human cancer
cell lines; Cervical (2008 and C13*), Colorectal (HT29 and HCT116) and Ovarian carcinoma
(A2780, A2780/CP and IGROV-1).

Highlights:
ꢀ
Single crystal X-ray analysis of compounds 1, 3 and various spectroscopic
techniques were used to establish the structure of the compounds.
ꢀ
The presence of intramolecular hydrogen bond (N
planarity of the molecule.
─H⋯O═C) compels
ꢀ
ꢀ
ꢀ
Hirshfeld surfaces analysis of compounds 1, 3 were performed to understand
hydrogen bonding and other weak interactions.
The compounds were screened for their In vitro cytotoxic activity against
seven human cancer cell lines.
The compounds screened for their cytotoxic activity in seven human cancer
cell lines exhibited promising anticancer activity

Declaration of interests
☐ The authors declare that they have no known competing financial interests or personal relationships
that could have appeared to influence the work reported in this paper.
☐The authors declare the following financial interests/personal relationships which may be considered
as potential competing interests:
The authors declare that they have no known competing financial interests or personal relationships that
could have appeared to influence the work reported in this paper.