Novel synthetic aromatic thiourea derivatives and investigations on their cytotoxic potential efficacy

Article abstract of DOI:10.1002/jhet.4145

Aromatic thiourea derivatives were synthesized by standard reactions from isothiocyanates in high yield. An efficient one-pot synthesis of primary amine with carbon disulfide involving the intermediate dithiocarbamate conversion using T₃P (propane phosphonic anhydride) to isothiocyanate is reported. Newly synthesized compounds showed excellent luminescence property. Cytotoxic investigation with different human cancer cell lines showed IC₅₀ values varying from of 17 to 40 μM for N-phenethyl-3,4-dihydroisoquinoline-2(1H)-carbothioamide (SCR172) among other molecules. Increase in Sub G1 population and increased depolarisation of mitochondria were evident in cells treated with SCR172, suggesting its role as a cancer therapeutic agent. This provides a lead structure for further synthetic modifications.

Full text of DOI:10.1002/jhet.4145

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Novel synthetic aromatic thiourea derivatives and investigations
on their cytotoxic potential efficacy
Ujjayinee Ray[a], Franklin John*[b], Sayeesh Pooppadi[b], Jinu George[b], Shivangi Sharma[a], Sathees C
Raghavan[a]
[a]
Miss. Ujjayinee Ray
Department of biochemistry
Indian Institute of Science
Bengaluru, India 560012
E-mail: ujjayineeray2@gmail.com
Miss. Shivangi Sharma
Department of biochemistry
Indian Institute of Science
Bengaluru, India 560012
E-mail: sharmashivangi1992@gmail.com
Prof. Dr. Sathees C Raghavan
Department of biochemistry
Indian Institute of Science
Bengaluru, India 560012
E-mail: sathees@iisc.ac.in
Prof. Dr. Franklin John
[b]
Department of chemistry
Sacred Heart College, Thevara
Kochi, India-682013
E-mail: franklinshc@gmail.com
Mr. Sayeesh pooppadi
Department of chemistry
Sacred Heart College, Thevara
Kochi, India-682013
E-mail: projectdbtsh@gmail.com
Prof. Dr. Jinu George
Department of chemistry
Sacred Heart College, Thevara
Kochi, India-682013
E-mail: jinujacobv@gmail.com
Abstract: Aromatic thiourea derivatives were synthesized by
standard reactions from isothiocyanates in high yield. An efficient
one- pot synthesis of primary amine with carbon disulfide involving
the intermediate dithiocarbamate conversion using T3P® (propane
phosphonic anhydride) to isothiocyanate is reported. Newly
synthesized compounds showed excellent luminescence property.
Cytotoxic investigation with different human cancer cell lines showed
IC50 values varying from of 17 to 40 μM for N-phenethyl-3,4-
dihydroisoquinoline-2(1H)-carbothioamide (SCR172) among other
molecules. Increase in Sub G1 population and increased
depolarisation of mitochondria was evident in cells treated with
SCR172, suggesting its role as cancer therapeutic agent. This
provides a lead structure for further synthetic modifications.
known that cells belonging to variety of human malignancies or
tumor growth have less ability to undergo apoptosis with regards
to at least some physiological stimuli.[2] In rapidly dividing cells
such as tumor cells, cytotoxicity has been attributed to DNA
modification.[3]
Cancer follows a variety of mutagenic processes which permit
cancer cells to gain extraordinary proliferation potential, self-
sufficiency in growth signals and resistance to both anti-
proliferative and apoptotic cues which would otherwise contain
their growth.[4,5] Proteins that regulate apoptosis can be
potential cancer drug targets.[6] Apoptosis inducers and cytotoxic
agents represent a wide spectrum of classical molecules having
anticancer ability.[6] Treating cells with cytotoxic agents can
cause decrease in cell viability or the cells can activate
apoptosis. Henceforth investigation on cytotoxic potential paves
way for an efficient method for the development of anticancer
agents.
Introduction
Search for novel anticancer drugs has led to the discovery of
synthetic imidazole derivatives (SCR1-SCR4) with anti-
carcinogenic activity and having less toxicity to the immune
system.[7] Palladium (II) and platinum (II) complexes exhibiting
Medicinal chemistry relies on novel molecules as bioactive
agents. Cancer is a global health problem and a leading cause
of death worldwide. Development of novel therapeutic agents
against cancer with lesser side effects is highly warranted. Low
bioavailability, less selectivity, acute toxicity and intra cellular
drug resistance demands the design and synthesis of novel
chemotherapeutic molecules for cancer treatment.[1] It is well
antitumor
along
with
luminescence
property
and
pharmacological activity were extensively studied earlier.[8]
Photoluminescence property of a molecular system is an added
advantage in bioanalytical applications. Schiffs base complexes
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and ZnO nanoparticles are known to exhibit photo physical and
anticancer properties.[9,10] Examples of other organic systems
are not so common but we elaborate the synthesis of a series of
compounds possessing both chemiluminescent and biological
properties.
Here we describe the synthesis, analytical characterization and
biological investigation of aromatic thiourea derivatives 3-benzyl-
4-imino-3,4-dihydroquinazoline-2-thione
chlorophenyl)-3,4-dihydroisoquinoline-2-carbothioamide
(SCR171), N-phenethyl-3,4-dihydroisoquinoline-2-
(SCR170),
N-(2-
Thiourea is a simple and well-known chemical entity and its
derivatives represent another important class of organic
molecules having broad spectrum of pharmacological activity.[11-
carbothioamide (SCR172) and 1-benzyl-3-(2-chlorophenyl)
thiourea (SCR173).
All the thioureas were synthesized starting from the
corresponding isothiocyanates by reaction with aromatic amines,
15]
Synthetic combinations of urea and thiourea connected with
benzothiazole moiety resulted in DNA topoisomerases and HIV
reverse transcriptase inhibitors.[16,17] Here we present the
synthesis, luminescence property investigation and cytotoxic
potential evaluation of thiourea derivatives (Figure 1). We
designed our synthetic strategy to obtain a thiourea entity
as
given
in
the
scheme
(Scheme
1).
2-
isothiocyanatobenzonitrile was reacted with benzyl amine at
room temperature to generate 2-cyanophenyl thiourea. The
latter is a thermally unstable intermediate which could be
cyclized by refluxing in ethanol leading to SCR170. (2-
isothiocyanatoethyl) benzene (2) was synthesized by a reported
procedure starting from phenethylamine.[23]
restricted in
a
heterocycle (SCR170) by reaction of
corresponding isothiocyanate (ITC) and benzyl amine followed
by cyclization. The next series of molecules SCR171 and
SCR172 was obtained by reaction of corresponding
isothiocyanate with tetrahydro isoquinoline resulting in one N-
end having a bulky group. SCR173 is another molecule
synthesized starting from the same ITC as SCR171. ITCs
represent an important class of precursor for the synthesis of
thioamides, sulphur containing heterocycles and thiourea
receptors.[18-20] Primary amines are used as the starting
molecule for the synthesis of ITCs. Phosphoethanolamines and
primary amines are key functional groups in biologically relevant
organic molecules. Many eukaryotic proteins are post
translationally modified with phosphoethanolamines at C-6
mannose sugar unit to anchor proteins.[21] Reduction of azides
and Gabriel’s phathalimide synthesis are examples of synthetic
procedures to obtain primary amines.[22] A two step one-pot
synthesis of primary amine with carbon disulfide via
dithiocarbamic acid salt through insitu desulfuration using
propane phosphonic anhydride (T3P®) was adopted.[23] T3P® is a
peptide coupling ‘green’ reagent which is stable and non-
toxic.[24,25]
Scheme 1. Synthetic Scheme
Synthesized compounds were analysed by IR, 1H-NMR and
HRMS spectra (supplementary data). All the compounds are
soluble in CHCl3, DMSO and ethyl acetate. The IR data
absorption showed strong band at 3300 cm-1 for NH group and
1430 cm-1 for C=S. Isothiocyanate group showed absorption
band at 2100 cm-1 (data not given). 13C NMR showed peaks at
around 훿 180 and 130 ppm which is characteristic for C=S.
Figure 1. Design strategy of thiourea derivatives
Results and Discussion
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Figure 4. Comparison of Cytotoxic effect of SCR172 in various cell lines. A-C.
Cytotoxicity induced by SCR172 was evaluated in various cancer cell lines,
Reh (A), K562 (B), Molt4 (C). Cells were treated with increasing
concentrations of the compound (10, 50, 100 and 250 μM) for 48 h and
cytotoxicity was evaluated. D. Table showing IC50 values (in μM) of SCR172 at
48 h.
Investigation of the luminescent properties exhibited by all four
thiourea compounds was carried out at room temperature within
350–600 nm wavelength range. This suggest that the
compounds fall in the category of luminescence materials
system having broad excitation bands (supplementary data).
The x-y coordinates of the chromaticity diagram belonging to the
emission spectra of samples SCR170, SCR171, SCR172 and
SCR173 were found (Figure 5) to be (0.182, 0.134), (0.202,
0.220), (0.185, 0.182), and (0.229, 0.315) respectively.[26]
Figure 2. Evaluation of cytotoxicity of thiourea derivatives (SCR170-173) in
leukemic cancer cell line, Nalm6. Cytotoxicity of the derivatives in Nalm6 at 48
h was evaluated at concentrations 0, 10, 50, 100 and 250 μM. Control cells
were treated with equivalent concentration of DMSO.
Cytotoxicity of synthesized compounds were analysed in human
cancer cell lines and the results revealed varying sensitivity.
Among the compounds, SCR172 showed good cytotoxic
potential in Nalm6 (Figure 2) and HeLa (Figure 3A). The IC50
value of SCR172 was found to be in the range of 17-40 μM
(Figure 3B).
Figure 5. Analysis of photoluminescence of thiourea derivatives. (x,y)
Chromaticity color coordinate diagram of compounds SCR170 (0.182, 0.134),
SCR171 (0.202, 0.220), SCR172 (0.185, 0.182), SCR173 (0.229, 0.315).
Since SCR172 showed good cytotoxicity in various cancer cell
lines, the effect on cell cycle progression was also investigated.
Results reveal a dose dependent increase in Sub G1 population
after treatment with SCR172 in Molt4 cell line (Figure 6), further
confirming increased cell death. A subsequent decrease in other
cell cycle phases was also observed. Further, with respect to
mitochondrial membrane potential, SCR172 treatment showed a
dose dependent increase in green cell population in Molt4
(Figure 7A, B) and in Nalm6 (Figure 7C, D) cells. This indicates
depolarisation of mitochondria post treatment with the
compound.
Figure 3. Effect of thiourea derivatives (SCR170-173) in HeLa cells. A.
Cytotoxicity of SCR170-173 in HeLa was evaluated at concentrations 0, 10,
50, 100 and 250 μM at 48 h time point. Control cells were treated with
equivalent concentration of DMSO. B. Table depicts IC50 of the derivatives in
HeLa and Nalm6 at 48 h.
SCR172 also showed significant cytotoxicity in leukemia cancer
cell lines, Reh, K562 and Molt4 in the range of 18-31 μM (Figure
4).
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lead structure opens up possibilities for further synthetic
modification for the facile synthesis of derivatives with improved
anticancer activity.
Figure 6. Cell cycle analysis of Molt4 cells after treatment with SCR172. A.
Analysis of cell cycle progression in Molt4 cells after treatment with SCR172 (0,
30, 60 μM). B. Bar graph represents percentage of cells in every stage of cell
cycle upon incubation with SCR172.
Experimental Section
Chemical synthesis
Solvents for moisture sensitive reactions were distilled and dried as per
standard procedures prior to use. All solvents used for silica gel flash
chromatography (FC) were distilled. Thin layer chromatography (TLC)
was carried out using Merck Silica Gel 60 F254 aluminium sheets. For
developing plates, KMnO4 was used. FC was performed on Merck Silica
Gel 60 (100-200 mesh). NMR spectra was recorded on Bruker AV-400
spectrometer at 298 K. Chemical shifts were referenced to the solvent
signal (CDCl3: δH = 7.26 δC = 77.0, DMSO-d6: δH = 2.5, δC = 39.5). ESI-
MS spectra were recorded on a Bruker Daltonics Esquire 3000 plus
instrument.
(2-Isothiocyanatoethyl)benzene (2)
Et3N (7.57 mL, 50 mmol) and CS2 (2.29 mL, 30 mmol) was added in one
portion to a solution of 2-phenylethanamine (1) (1.27 mL, 10 mmol) in
DCM (10 mL) and poured to a 50 mL two-neck round-bottom flask fitted a
rubber septum, and thermometer. This was secured from moisture using
a syringe filled with CaCl2. Solution was stirred for 1 h at r.t. The
temperature of the reaction mixture was adjusted to 4 °C and T3P® (5.39
mL, 18 mmol) was added in a span of 5 min in three portions. Thereafter,
the solution was allowed to reach rt. and was stirred for 2 h. The reaction
mixture was stirred with water (10 mL) for 30 min and DCM (50 mL) was
added. Then the organic layer was separated and washed successively
with water (2 × 5 mL), 1 M HCl (2 × 5 mL), saturated NaHCO3 (2 × 5 mL),
and saturated NaCl solution (5 mL). The solution was dried over
anhydrous Na2SO4. The crude product was purified by FC (Hexane:Ethyl
acetate, 1:1) to get a colourless gum (1.36 g, 8 mmol, 83%)
1H NMR (400 MHz, DMSO-d6): δ = 7.37–7.23 (m, 5H), 3.92 (t, 2H, J =
8.0 Hz), 2.97 (t, 2H, J = 8.0 Hz) 13C NMR (100 MHz, DMSO-d6): δ =
137.6 (NCS), 128.8, 128.4, 126.8, 126.3, 45.9, 35.3. ESI-MS (M+H)+
calculated for C9H10NS m/z 164.0534, found m/z 164.0532
Figure 7. Effect of SCR172 on loss in mitochondrial membrane potential A.
JC-1 assay was carried out for detecting loss in mitochondrial membrane
potential of Molt4 cells following treatment with SCR172 in a dose-dependent
manner (0, 30 and 60 μM). 2, 4-DNP (4 mM) was used as positive control. B.
Bar graph shows the percentage of green cells experiencing loss in membrane
potential compared to that of red cells in Molt4 cells. C. JC-1 assay in Nalm6
treated with SCR172 (0, 20 and 40 μM). D. Percentage quantitation of Nalm6
cells experiencing loss in membrane potential is shown.
3-benzyl-4-imino-3,4-dihydroquinazoline-2(1H)-thione SCR170
A solution of 2-isothiocyanatobenzonitrile (0.1g, 0.624 mmol) in DCM (20
mL) was mixed with benzyl amine (0.0681 mL, 0.624 mmol) and stirred
at rt for 10 min. After the disappearance of 2-isothiocyanatobenzonitrile
(checked by TLC), solvent was removed under vacuum. The crude
product obtained was refluxed in ethanol for 3h. Solvent was evaporated
and purified by FC (Hexane/Ethyl acetate, 1:1) to give SCR 170 (0.1169g,
0.4372 mmol, 70%) as a pale yellow powder (Rf = 0.7, Hexane/Ethyl
acetate, 1:1). 1H NMR (400 MHz, DMSO-d6) δ 12.3 (s, 1H), 9.46 (s, 1H),
Conclusions
Thus, in the current study we report the chemistry,
photoluminescence properties and cytotoxicity of aromatic
thiourea derivatives, synthesized by a one-pot reaction starting
from the corresponding isothiocyanates. Among the synthesized
compounds, SCR172 showed improved cytotoxic potential in
various cancer cell lines with IC50 around 17-40 μM. The
compounds also exhibited good luminescence activity at room
temperature. Increase in Sub G1 population and depolarised
mitochondrial membrane potential also confirmed increased cell
death with SCR172 in a dose-dependent manner. Thus, this
8.11-8.09 (m, 1H), 7.61-7.57 (m, 1H), 7.33-7.19 (m, 7H), 5.88 (s, 2H).13
C
NMR (100 MHz, DMSO-d6) δ 180.3, 138.5, 133.3, 132.1, 131.7, 131.4,
129.4, 54.9. ESI-MS (M+H)+ calculated for C15H34N3S m/z 268.0908,
found m/z 268.0832.
N-(2-chlorophenyl)-3,4-dihydroisoquinoline-2(1H)-carbothioamide
SCR171
1,2,3,4-tetrahydroisoquinoline (0.6258 mL, 5 mmol) was added drop wise
to a solution of 1-chloro-2-isothiocyanatobenzene (0.659 mL, 5 mmol) in
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acetonitrile (10 mL), and stirred at room temperature for 10 min. After
the disappearance of 1-chloro-2-isothiocyanatobenzene (checked by
TLC), solvent was evaporated off. Residue was recrystallized using
ethanol to give SCR171 (0.33 g, 1.10 mmol, 70%) as white crystals (Rf =
0.8, Hexane/ethyl acetate, 1:1).1H NMR (400 MHz, CDCl3) δ 7.85-7.82
(m, 1H), 7.43-7.41 (m, 1H), 7.30-7.18 (m, 6H), 75.03 (s, 2H), 4.08 (t, 2H,
J = 4.0 Hz), 3.03 (t, 2H, J = 4.0 Hz).13C NMR (100 MHz,CDCl3) δ 181.7,
129.5, 127.9, 127.4, 127.0, 126.9, 126.5, 126.1, 126.0, 50.7, 46.8, 29.2.
ESI-MS (M+H)+ calculated for C16H16ClN2S m/z, 303.0729, found m/z
303.0723.
Cell cycle analysis
Cell cycle analysis was performed as described earlier.30,31 Cells were
treated with SCR172 (30 and 60μM for Molt4) for 48 h. After harvesting
cells were washed with 1X PBS and fixed in 70% ethanol overnight at -
20oC. RNaseA treatment was given overnight at 37oC. Finally, cells were
stained with Propidium Iodide and analyzed using CytoFLEX
flowcytometer, Beckman Coulter (Brea, California, United States). 10,000
cells were acquired minimum and data analysis was carried out by using
CytExpert software.
N-phenethyl-3,4-dihydroisoquinoline-2(1H)-carbothioamide SCR172
JC-1 assay
To a solution of (2-Isothiocyanatoethyl)benzene (0.82 g, 5 mmol) in
Acetonitrile (10 mL), 1,2,3,4-tetrahydroisoquinoline (0.62 mL, 5 mmol)
was added drop wise and stirred at rt for 10 min. Solvent was evaporated
after the disappearance of (2-Isothiocyanatoethyl)benzene (checked by
TLC). Residue was recrystallized from ethanol to yield SCR172 (1.1 g, 3
mmol, 75%), as brown crystals (Rf = 0.8, Hexane/Ethyl acetate, 1:1).1H
NMR (400 MHz, DMSO-d6) δ 7.81 (m, 1H), 7.40– 7.02 (m, 8H), 4.93 (s,
2H), 3.94 (t, J = 4.0 Hz, 2H), 3.75 (m, 2H), 2.92-2.85 (m, 4H).13C NMR
(100 MHz,DMSO-d6) δ 168.1, 161.6, 159.0, 151.0, 154.4, 147.8, 143.6,
136.3, 128.2, 127.8, 123.5, 121.0, 119.6, 119.4, 106.6, 47.3, 29.4, 27.4
ESI-MS (M+H)+calculated for C18H21N2S m/z 297.1425, found m/z
297.1428.
In order to identify changes in mitochondrial membrane potential,
treatment of the cells were carried out with increasing concentration of
SCR172 (20 and 40 μM for Nalm6; 30 and 60 μM for Molt4) for 48 h and
subjected to JC-1 staining assay.27 Cells were incubated in JC-1 dye
(5,50,6,60-tetrachloro-1,1,3,30-
tetraethylbenzimidazolcarbocyanamideiodide (Calbiochem, USA) (1 μM)
at 37oC for 20 min, with intermittent mixing as described earlier.28 2, 4-
Dinitrophenol (4 mM) was used as positive control. After 1X PBS wash,
samples were acquired using CytoFLEX flowcytometer. Mitochondrial
depolarization, indicated by a decrease in the red/green fluorescence
intensity ratio, was represented in bar diagrams prepared using
GraphPad Prism.
1-benzyl-3-(2-chlorophenyl)thiourea SCR173
Photoluminescence
A solution of 1-chloro-2-isothiocyanatobenzene (0.66 mL, 5 mmol) in
acetonitrile (10 mL), was mixed with benzyl amine (0.5 mL, 5mmol) and
stirred at rt for 10 min. After the disappearance of 1-chloro-2-
isothiocyanatobenzene (checked by TLC), solvent was evaporated off
and the residue was recrystallized from ethanol to give SCR173 (1.05g,
3mmol, 75%) as white crystals (Rf = 0.7, Hexane/Ethyl acetate, 1:1).
Luminescent spectrum of thiourea derivatives was obtained at room
temperature. The excitation spectra (supplementary data) from
wavelength of 350–600 nm indicates that the synthesized compounds
belong to the luminescence materials system with broad excitation bands.
Commission International deI’Eclairage (1931) system for the emission
spectrum was used to find colour coordinate (x, y) and colour purity of
the compounds.[26]
1H NMR (400 MHz, CDCl3) δ 7.63-7.21 (m, 9H), 4.89 (d, J = 8.0 Hz,
2H).13C NMR (100 MHz, CDCl3) δ 181.1, 128.9, 128.4, 128.1, 127.9,
127.7, 127.1, 49.6. ESI-MS (M+H)+calculated for C14H14ClN2S m/z
277.0566, found m/z, 277.0570.
Acknowledgements
The reported work was financially supported by grants from
KSCSTE (564/2017/KSCSTE, 5130/SRSPS), Govt of Kerala to
JG, and CEFIPRA (IFC/5203-4/2015/131), DAE (21/01/2016-
Cell culture
Nalm6 and Reh (B-cell precursor leukemia), K562 (myelogenous
leukemia) and Molt4 (T-cell acute lymphoblastic leukemia) cells were
cultured in RPMI 1640 containing 5% FBS, 100 μg/ml of antibiotics
Penicillin G and Streptomycin. Human cervical cancer cell line HeLa was
cultured in DMEM containing 5% FBS and PenStrep. Cells were
incubated in a humidified atmosphere at 37oC containing 5% CO2 as
described before.27
BRNS/35074)
and
IISc-DBT
partnership
programme
[DBT/BF/PR/INS/2011-12/IISc] to SCR. UR, FJ and SP
performed the experiments. FJ, UR and SCR wrote the
manuscript and analysed the data. UR was supported by Senior
Research fellowship (SRF) from CSIR, India.
Keywords: Thiourea, Cytotoxicity, Photoluminescence, Cell
Cytotoxicity
cycle, Cancer therapeutics, Mitochondrial membrane potential
Trypan Blue dye exclusion assay was used to evaluate effect of the
compounds in cancer cell lines as described earlier.28,29 25000 cells/ml
were seeded in tissue culture grade 24 well plates and treated with
increasing concentrations of the compounds (10, 50,100 and 250 μM).
Equivalent amount of DMSO was added to the control. Cytotoxicity was
evaluated at 24 h and 48 h for suspension cell lines and 48 h for
adherent cell line HeLa. IC50 values were calculated for each compound
using GraphPad Prism. Cells were mixed with same volume of 0.4%
Trypan blue (Sigma Chemical Co., St Louis, MO, USA) and counted
using haemocytometer. Experiments were repeated at least three times
and error bars were plotted using GraphPad Prism.
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equivalent concentration of DMSO. B. Table depicts IC50 of the derivatives in
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Figure 4. Comparison of Cytotoxic effect of SCR172 in various cell lines. A-C.
Cytotoxicity induced by SCR172 was evaluated in various cancer cell lines,
Reh (A), K562 (B), Molt4 (C). Cells were treated with increasing
concentrations of the compound (10, 50, 100 and 250 μM) for 48 h and
cytotoxicity was evaluated. D. Table showing IC50 values (in μM) of SCR172 at
48 h.
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Figure 5. Analysis of photoluminescence of thiourea derivatives. (x,y)
Chromaticity color coordinate diagram of compounds SCR170 (0.182, 0.134),
SCR171 (0.202, 0.220), SCR172 (0.185, 0.182), SCR173 (0.229, 0.315).
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Figure 6. Cell cycle analysis of Molt4 cells after treatment with SCR172. A.
Analysis of cell cycle progression in Molt4 cells after treatment with SCR172 (0,
30, 60 μM). B. Bar graph represents percentage of cells in every stage of cell
cycle upon incubation with SCR172.
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Figure 7. Effect of SCR172 on loss in mitochondrial membrane potential A.
JC-1 assay was carried out for detecting loss in mitochondrial membrane
potential of Molt4 cells following treatment with SCR172 in a dose-dependent
manner (0, 30 and 60 μM). 2, 4-DNP (4 mM) was used as positive control. B.
Bar graph shows the percentage of green cells experiencing loss in membrane
potential compared to that of red cells in Molt4 cells. C. JC-1 assay in Nalm6
treated with SCR172 (0, 20 and 40 μM). D. Percentage quantitation of Nalm6
cells experiencing loss in membrane potential is shown.
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Figure legends
Figure 1. Design strategy of thiourea derivatives
Scheme 1. Synthetic Scheme
Figure 2. Evaluation of cytotoxicity of thiourea derivatives (SCR170-173) in
leukemic cancer cell line, Nalm6. Cytotoxicity of the derivatives in Nalm6 at 48
h was evaluated at concentrations 0, 10, 50, 100 and 250 μM. Control cells
were treated with equivalent concentration of DMSO.
Figure 3. Effect of thiourea derivatives (SCR170-173) in HeLa cells. A.
Cytotoxicity of SCR170-173 in HeLa was evaluated at concentrations 0, 10,
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