Деякі фармакологічні властивості 4-[3-(5-бромо-2-гідроксифеніл)-5-феніл-3,4-дигідропіразол-2-іл]-5Н-тіазол-2-ону
A series of 3,5-diaryl pyrazolyl thiazolinones were designed and synthesized as potential biologically active compounds. The study of anticancer activity of 4-[3-(5-bromo-2-hydroxyphenyl)-5-phenyl-3,4-dihydropyrazol-2-yl]-5H-thiazol-2-one (1) revealed its high antiproliferative activity against a pa...
Збережено в:
| Дата: | 2020 |
|---|---|
| Автор: | |
| Формат: | Стаття |
| Мова: | Англійська |
| Опубліковано: |
V.P. Kukhar Institute of Bioorganic Chemistry and Petrochemistry of the National Academy of Sciences of Ukraine
2020
|
| Теми: | |
| Онлайн доступ: | https://bioorganica.com.ua/index.php/journal/article/view/40 |
| Теги: |
Додати тег
Немає тегів, Будьте першим, хто поставить тег для цього запису!
|
| Назва журналу: | Ukrainica Bioorganica Acta |
| Завантажити файл: |  |
Репозитарії
Ukrainica Bioorganica Acta| _version_ | 1871193576981921792 |
|---|---|
| author | Kryshchyshyn-Dylevych, Anna |
| author_facet | Kryshchyshyn-Dylevych, Anna |
| author_institution_txt_mv | [
{
"author": "Anna Kryshchyshyn-Dylevych",
"institution": "Danylo Halytsky Lviv National Medical University, 69 Pekarska, Lviv, 79010, Ukraine"
}
] |
| author_sort | Kryshchyshyn-Dylevych, Anna |
| baseUrl_str | https://bioorganica.com.ua/index.php/journal/oai |
| collection | OJS |
| datestamp_date | 2026-07-19T14:56:53Z |
| description | A series of 3,5-diaryl pyrazolyl thiazolinones were designed and synthesized as potential biologically active compounds. The study of anticancer activity of 4-[3-(5-bromo-2-hydroxyphenyl)-5-phenyl-3,4-dihydropyrazol-2-yl]-5H-thiazol-2-one (1) revealed its high antiproliferative activity against a panel of cancer cells with the lowest growth inhibition concentration (GI50) towards leukemic cell line SR (0.0351 µМ) and ovarian cancer cell line OVCAR-3 (0.248 µМ). It was also found that pyrazolyl thiazolinone 1 inhibited growth of Trypanosoma brucei brucei by 98,8% at a concentration of 10 µg/mL. The in-depth cytotoxicity study of compound 1 on human hepatocellular carcinoma HepG2 cells and non-tumorigenic murine fibroblast Balb/c 3T3 in MTT, NRU, TPC and LDH assays showed that normal cells were less sensitive to compound 1 than the cancer cells; its action had led to a disintegration of the cell membrane, inhibition of mitochondrial and lysosomal activity, and proliferation of cancer cells. The highest selectivity were detected in the LDH assay |
| doi_str_mv | 10.15407/bioorganica2020.02.041 |
| first_indexed | 2025-07-17T12:19:41Z |
| format | Article |
| fulltext |
ISSN 1814-9758. Ukr. Bioorg. Acta, 2020, Vol. 15, N 2
UDC 615.012.1:547.789/.793
DOI: https://doi.org/10.15407/bioorganica2020.02.041
41
RESEARCH ARTICLE
Some pharmacological properties of 4-[3-(5-bromo-2-hydroxyphenyl)-5-
phenyl-3,4-dihydropyrazol-2-yl]-5H-thiazol-2-one
Anna Kryshchyshyn-Dylevych*
Danylo Halytsky Lviv National Medical University, 69 Pekarska, Lviv, 79010, Ukraine
Abstract: A series of 3,5-diaryl pyrazolyl thiazolinones were designed and synthesized as potential biologically active compounds. The
study of anticancer activity of 4-[3-(5-bromo-2-hydroxyphenyl)-5-phenyl-3,4-dihydropyrazol-2-yl]-5H-thiazol-2-one (1) revealed its high
antiproliferative activity against a panel of cancer cells with the lowest growth inhibition concentration (GI50) towards leukemic cell line
SR (0.0351 µМ) and ovarian cancer cell line OVCAR-3 (0.248 µМ). It was also found that pyrazolyl thiazolinone 1 inhibited growth of
Trypanosoma brucei brucei by 98,8% at a concentration of 10 µg/mL. The in-depth cytotoxicity study of compound 1 on human
hepatocellular carcinoma HepG2 cells and non-tumorigenic murine fibroblast Balb/c 3T3 in MTT, NRU, TPC and LDH assays showed
that normal cells were less sensitive to compound 1 than the cancer cells; its action had led to a disintegration of the cell membrane,
inhibition of mitochondrial and lysosomal activity, and proliferation of cancer cells. The highest selectivity were detected in the LDH
assay.
Keywords: pyrazolyl thiazolinone hybrids; antitumor activity; antitrypanosomal activity; cytotoxicity.
Introduction
The hybrid pharmacophore approach has been used to
develop a drug-like molecules with anticancer properties
[1]. Pyrazole and thiazole/thiazolidinone cycles are
considered as favored scaffolds and have been used to
design novel drug-like molecules possessing antiprolife-
rative activity [2-3]. A combination of the above
heterocycles in one molecule is likely to provide more
efficient pyrazolyl-thiazole/thiazolidone conjugates via
synergistic anticancer effects. For example, a 2-amino-
iminothiazolidinone derivative with a pyrazole moiety at
position 5 inhibited necroptosis [4], pyrazolyl-4-thiazolid-
inones exhibited dose-dependent cytotoxic effect in human
breast cancer cells (MCF-7 line) [5], a series of 2-(5-aryl-3-
phenyl-4,5-dihydro-1H-pyrazol-1-yl)-1,3-thiazol-4(5H)ones
inhibited growth of leukemia cell lines and non-small cell
Received:
Revised:
Accepted:
Published online:
02.11.2020
10.11.2020
20.11.2020
30.12.2020
Corresponding author. Tel.: +380-32-275-5966;
e-mail: kryshchyshyn.a@gmail.com (A. P. Kryshchyshyn-Dylevych)
ORCID: 0000-0002-8713-7020
lung cancer cell lines [6]. Cytotoxicity of pyrazole
derivatives includes inhibition of the topoisomerase I and II
activity for pyrazoloacridine [7] or inhibition of the Janus-
activated kinase (JAK1/2) for the Ruxolitinib [8]. The
pyrazolyl thiazol/thiazolidinone-based hybrids as biologi-
cally active compounds have been studied for anti-parasitic
activity [9-10]. For example, some of the 5-(3,5-diaryl-4,5-
dihydropyrazol-1-ylmethylene)-2-thioxothiazolidin-4-ones
showed IC50 level of activity within 0.6-0.7 µM in vitro
assay towards Trypanosoma brucei gambiense [11];
5-[5-aryl-3-naphthalen-2-yl-4,5-dihydropyrazol-1-yl]-thia-
zolidine-2,4-dione and 2-{5-[5-aryl-3-naphthalen-2-yl-4,5-
dihydropyrazol-1-yl]-2,4-dioxothiazolidin-3-yl}-N-arylace-
tamides inhibited growth of the Trypanosoma brucei brucei
and Trypanosoma brucei gambiense at micromolar
concentrations [12]. Thiazole/thiazolidinone phenylindole/
imidazothiadiazoles comprise another highly active class of
antitrypanosomals [13]. The combination of different
pharmacophores in one molecule may lead to novel drug-
like molecules that will exhibit biological activity. Thus, the
above mentioned polypharmacological [14] and hybrid-
pharmacophore approaches [1] inspire the design and
synthesis of pyrazolyl-thiazolinones as potential anticancer
and antiparasitic agents.
© Kryshchyshyn-Dylevych A. P. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits
unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Ukrainica Bioorganica Acta
www.bi oorgan ica .org .ua
ISSN 1814-9758. Ukr. Bioorg. Acta, 2020, Vol. 15, N 2
42
S
NH
S
O
N
H
N
OH
Br
Ph
EtOH
N
N
Ph HO
Br
S
N
O
AcONa
AcOH
1
R = H (a); 4-MeO (b); 3-NO2 (c)
O
R
N
N
Ph HO
Br
S
N
O
2
R
Scheme 1. Synthesis of a pyrazolyl-thiazolinone 1 and its 5-arylidene derivatives 2a-c.
Results and Discussion
A 4-[3-(5-bromo-2-hydroxyphenyl)-5-phenyl-3,4-dihyd-
ropyrazol-2-yl]-5H-thiazol-2-one (1) was synthesized in the
reaction of 3,5-diarylpyrazoline with 4-thioxo-2-
thiazolidinone (isorhodanine) in the ethanol by known
approach [6]. 5-arylidene derivatives 2a-c were synthesized
in high yields by the Knoevenagel reaction with correspon-
ding aldehydes (Scheme 1). 4-[3-(5-Bromo-2-hydroxyphe-
nyl)-5-phenyl-3,4-dihydro-pyrazol-2-yl]-5H-thiazol-2-one
(1) was selected by National Cancer Institute (NCI,
Bethesda, USA) Developmental Therapeutic Program
(DTP) for anticancer screening at one dose assay (10-5 M)
using a panel of 59 cancer cell lines that represented
different types of cancer (leukemia, melanoma, lung, colon,
CNS, ovarian, renal, prostate and breast cancers). The
anticancer activity of compound 1 showed the mean growth
percent for the whole cancer cell panel and it was about
25% [15-17]. Moreover, the tested pyrazolyl thiazolinone 1
demonstrated the cytotoxic effects to five leukemic cell
lines (CCRF-CEM, HL-60(TB), MOLT-4, RPMI-8226,
SR), a non-small cell lung cancer cell line (HOP-92), a CNS
cancer cell line (SF-295), and an ovarian cancer cell line
(IGROV1). The high growth inhibition rates of compound 1
justified the in-depth screening at a range of concentrations
using 55 cell lines. The percentage of growth was evaluated
spectrophotometrically versus control that were not treated
with tested compound after 48 h of exposure. The SRB
protein assay was used to estimate cells’ viability or growth.
Three dose-response parameters for antitumor activity were
found for each cell line: GI50 – molar concentration of the
compound that inhibits 50% net cell growth; TGI – molar
concentration of the compound leading to the total
inhibition; and LC50 – molar concentration of the compound
leading to 50% net cell death. The high antiproliferative
activity of compound 1 against the majority of cell is
reported in Table 1. The lowest inhibitory concentrations
for compound 1 were observed against the leukemia panel.
The most sensitive leukemic cell line was the SR line (GI50
= 0.0351 μM). It inhibited growth of CCRF-CEM, HL-60,
MOLT-4 and SR leukemia lines at submicromolar concen-
trations (TGI). The high cytotoxic activity of 1 was found
against non-small cell lung cancer line NCI-H322M and
EKVX as well as colon cancer line HCT-116, ovarian
cancer OVCAR-3, and breast cancer MCF7 lines.
The antitrypanosomal activity of pyrazolyl-thiazolinone
1 on against T. brucei brucei was investigated in vivo by the
polypharmacological approach. A combination of pyrazo-
line and thiazolinone cycles in one molecule did not only
contribute to the anticancer properties, but also increased
the antiparasitic activity. It was found that 4-[3-(5-bromo-2-
hydroxyphenyl)-5-phenyl-3,4-dihyd-ropyrazol-2-yl]-5H-
thiazol-2-one (1) inhibited growth of T. brucei brucei by
98,8% at a concentration of 10 µg/mL. Lowering
concentration to 1 µg/mL led to sharp decrease in the
inhibition activity (11,87 % of inhibition).
The biological assays demonstrated high antitumor and
antitrypanosomal activities of pyrazolyl thiazolinone 1.
High activities inspired further in-depth studies of 1 against
human hepatocellular carcinoma HepG2 cells and non-
tumorigenic murine fibroblast Balb/c 3T3. Four different
biochemical endpoints were measured: mitochondrial
activity, lysosomal activity, total protein content, and
cellular membrane integrity in MTT assay, NRU assay,
TPC assay and LDH assay respectively (after 24, 48 and
72 h exposition).
The cells viability after the exposure to tested compound
was found to be a time-, a concentration- and a cell line-
dependent (Figure 1). Human hepatoma HepG2 cells were
more sensitive to the compound 1 compared to normal
murine Balb/c 3T3 fibroblast cells. The first sign of
anticancer activity of compound 1 was seen at the lowest
concentrations after 24 h of exposure (LDH assay).
Moreover, the toxic effect for normal fibroblasts cell line in
the same assay was observed at a concentration of 0.9 µM
(CC20) after 48 h of exposure (Figure 1). The calculated
cytotoxic concentrations values (CC20, CC50, and CC80) for
the cancer HepG2 cell line and for the non-tumorigenic
immortalized Balb/c 3T3 cell line are shown in Table 2. It is
worth to note that the lowest cytotoxic concentrations
(CC20) that inhibits 20% of cell growth were calculated for
all tests regardless of the exposure time. CC50 values
indicated that the first step of anticancer action of
compound 1 was the first disintegration of cellular
membranes leading to the inhibition of mitochondrial
activity with further inhibition of proliferation and
lysosomal activity. It is noteworthy that compound 1
exhibits low toxicity against normal murine embryonic
fibroblast cell line (Balb/c 3T3). Its toxic action led to dis-
A. P. Kryshchyshyn-Dylevych
43
Table 1. Influence of the 4-[3-(5-bromo-2-hydroxyphenyl)-5-phenyl-3,4-dihydropyrazol-2-yl]-5H-thiazol-2-one (1) on the
growth of the most sensitive tumor cell lines in in vitro test (10-4-10-8М).
Cancer cell lines GI50, µМ TGI, µМ LC50, µМ
Leukemia
CCRF-CEM
HL-60
K-562
MOLT-4
SR
0.339
0.306
0.959
0.397
0.0351
2.70
0.923
>100.0
17.7
0.282
>100.0
>100.0
>100.0
>100.0
>100.0
Non-small cell lung
cancer
A549/ATCC
EKVX
NCI-H23
NCI-H322M
NCI-H460
6.55
0.382
0.819
0.306
0.619
>100.0
>100.0
>100.0
15.3
>100.0
>100.0
>100.0
>100.0
>100.0
>100.0
Colon cancer
HCC-2998
HCT-116
HCT-15
SW-620
1.94
0.302
0.767
0.694
8.58
>100.0
>100.0
>100.0
>100.0
>100.0
>100.0
>100.0
CNS cancer
SF-268
SF-295
0.805
0.636
>100.0
>100.0
>100.0
>100.0
Melanoma
MALME-3M
M14
MDA-MB-435
SK-MEL-5
0.843
0.907
1.92
1.18
22.3
28.4
>100.0
20.8
>100.0
>100.0
>100.0
>100.0
Ovarian cancer
OVCAR-3
OVCAR-4
NCI/ADR-RES
SK-OV-3
0.248
0.464
0.830
1.84
0.682
>100.0
20.7
36.0
>100.0
>100.0
>100.0
>100.0
Renal cancer
ACHN
CAKI-1
TK-10
1.44
0.723
1.62
>100.0
4.06
>100.0
>100.0
>100.0
>100.0
Prostate cancer DU-145 2.24 >100.0 >100.0
Breast cancer
MCF7
T-47D
MDA-MB-468
0.301
0.377
0.631
-
>100.0
67.6
>100.0
>100.0
>100.0
integration of the cell membrane, inhibition of mitochondri-
al and lysosomal activity of cancer cells.
The selectivity indexes (SI), which indicates the
cytotoxic selectivity (i.e. drug safety) were calculated for
HepG2 cell line against corresponding normal cell line
(Figure 2). Higher values of SI indicate higher anticancer
specificity. Compounds with SI above 3.0 are considered
highly selective [18]. The highest values of SI were
detected in the LDH assay for compound 1. The SI
increased from 167 to 4975 with increase of the exposure
time from 24 h to 72 h whereas SI values for the cisplatin
increased only from 13 to 1429 (Figure 2).
ISSN 1814-9758. Ukr. Bioorg. Acta, 2020, Vol. 15, N 2
44
Concentration [M]
M
T
T
r
e
d
u
c
ti
o
n
[
%
]
10- 6 10- 5 10 - 4 10- 3 10- 2 10 - 1 100 101 102 103
0
25
50
75
100
Concentration [M]
M
T
T
r
e
d
u
c
ti
o
n
[
%
]
10 - 6 10 - 5 10- 4 10- 3 10- 2 10- 1 100 101 102 103
0
25
50
75
100
Concentration [M]
N
e
u
tr
a
l
re
d
u
p
ta
k
e
[
%
]
10- 6 10 - 5 10- 4 10- 3 10- 2 10- 1 100 101 102 103
0
25
50
75
100
Concentration [M]
N
e
u
tr
a
l
re
d
u
p
ta
k
e
[
%
]
10 - 6 10 - 5 10- 4 10- 3 10- 2 10- 1 100 101 102 103
0
25
50
75
100
Concentration [M]
T
o
ta
l
c
e
llu
la
r
p
ro
te
in
[
%
]
10- 6 10 - 5 10- 4 10- 3 10- 2 10- 1 100 101 102 103
0
25
50
75
100
Concentration [M]
T
o
ta
l
c
e
llu
la
r
p
ro
te
in
[
%
]
10 - 6 10 - 5 10- 4 10- 3 10- 2 10- 1 100 101 102 103
0
25
50
75
100
Concentration [M]
L
D
H
r
e
le
a
s
e
%
10- 6 10 - 5 10- 4 10 - 3 10- 2 10 - 1 100 101 102 103
0
25
50
75
100
24 h 48 h 72 h
Concentration [M]
L
D
H
r
e
le
a
s
e
%
10- 6 10 - 5 10- 4 10 - 3 10- 2 10 - 1 100 101 102 103
0
25
50
75
100
24 h 48 h 72 h
HepG2 Balb/c 3T3
Figure 1. Cytotoxic concentrations (CC20; CC50; CC80) (µM) calculated for compound 1 in normal and cancer cells.
A. P. Kryshchyshyn-Dylevych
45
Conclusions
High anticancer activity of 4-[3-(5-bromo-2-hydro-
xyphenyl)-5-phenyl-3,4-dihydropyrazol-2-yl]-5H-thiazol-2-
one (1) against a series of cancer cell lines with increased
selectivity against leukemic cell lines had been
demonstrated. (3,4-Dihydropyrazol-2-yl)-5H-thiazol-2-one
(1) showed a good antitrypanosomal activity in the in vitro
experiment against T. brucei brucei. The dual anticancer
and antitrypanosomal activity along with high selectivity
indices as well as low cytotoxicity would justify further in-
depth studies of compound 1 as a perpective drug candidate.
Table 2. The values of cytotoxic concentrations (CC20; CC50; CC80) (µM) calculated for compound 1 in MTT, NRU, TPC
and LDH assays.
Cell line Method Time (h) CC20 CC50 CC80
*
Balb/c 3T3
MTT
24 0.9±0.3 8.9±0.9
48 0.8±0.2 5.8±0.6 52.1±5.2
72 0.1±0.1 2.5±0.8 7.4±0.5
NRU
24 2.4±1.1 - -
48 0.2±0.1 7.1±1.5
72 0.05±0.002 2.1±1.0 7.1±0.3
TPC
24 2.3±1.0 - -
48 0.6±0.3 6.0±0.5
72 0.006±0.002 2.8±0.7 6.5±0.9
LDH
24 - - -
48 9.2±0.6 - -
72 0.5±0.1 19.9±2.6 92.9±2.5
HepG2
MTT
24 0.9±0.09 6.9±0.5 -
48 0.1±0.04 5.5±0.2 42.1±2.5
72 0.03±0.01 0.44±0.02 0.97±0.04
NRU
24 0.07±0.02 34.2±1.7 -
48 0.04±0.01 5.9±0.7 -
72 0.001±0.0003 1.3±0.1 6.6±0.1
TPC
24 0.7±0.2 95.5±0.5 -
48 0.05±0.02 5.4±0.4 -
72 0.001±0.0002 0.9±0.04 6.2±0.1
LDH
24 0.06±0.01 0.6±0.02 -
48 0.006±0.001 0.5±0.07 47.9±4.9
72 0.0003±0.0001 0.004±0.001 0.3±0.07
* CC20, CC50 and CC80 (µM) represents the concentrations of compound 1 that is required for 20%, 50% and 80 % inhibition using the MTT, NRU, TPC,
and LDH assays. Data are expressed as the mean ± SEM and were calculated from the dose response curves of at least three independent experiments.
ISSN 1814-9758. Ukr. Bioorg. Acta, 2020, Vol. 15, N 2
46
Figure 2. Selectivity indices (SI) calculated for compound 1 and cisplatin against HepG2 cells.
Experimental section
Chemistry
Commercial reagents were purchased from Merck and
used without purification. Melting points were measured in
open capillary tubes on a BŰCHI B-545 melting point
apparatus and are uncorrected. The elemental analyses were
performed using the Perkin-Elmer 2400 CHN analyzer. The
1H NMR spectra were recorded on Varian Gemini (1H at
400 and 13C at 100 MHz) instrument in DMSO‑d6 using
tetramethylsilane (TMS) as an internal standard.
The starting 3,5-diaryl-4,5-dihydropyrazole was synthe-
sized according to known method from chalcone [19].
4-[3-(5-Bromo-2-hydroxyphenyl)-5-phenyl-3,4-dihydro-
pyrazol-2-yl]-5H-thiazol-2-one (1).
A mixture of 4-thioxo-2-thiazolidinone (0.01 mol) and
4-bromo-2-(3-phenyl-4,5-dihydro-1H-pyrazol-5-yl)phenol
(0.01 mol) was refluxed in 100 ml of ethanol for 1.5 h.
After cooling the reaction mixture to room temperature,
formed precipitate was filtered off, washed with methanol
and recrystallized. Yield: 65%; pale yellow solid; mp 232-
234 °C, (DMF-EtOH 1:2). 1H NMR (400 MHz, CDCl3) δ
7.86 (d, J 8.2 Hz, 2Н, Ar), 7.47-7.56 (m, 3Н, Ar), 7.24 (dd,
J 2.7, 8.2 Hz, 1Н, Ar), 7.10 (d, J 2.7 Hz, 1Н, Ar), 6.83 (d,
J 8.2 Hz, 1Н, Ar), 5.75 (dd, J 3.3, 20.0 Hz, 1Н, С5H-
pyraz.), 4.87 (d, J 16.8 Hz, 1Н, С4H-thiaz.), 4.65 (d, J 16.8
Hz, 1Н, С4H-thiaz.), 4.00 (dd, J 11.1, 20.0 Hz, 1Н, С4H-
pyraz.), 3.25 (dd, J 3.3, 20.0 Hz, 1Н, С4H-pyraz.). Anal.
Calcd. for C18H14BrN3O2S: C, 51.93; H, 3.39; N, 10.09.
Found: C, 52.05; H, 3.45; N, 9.93.
General procedure for synthesis of compounds 2a-c
The equimolar amounts of 4-[3-(5-bromo-2-
hydroxyphenyl)-5-phenyl-3,4-dihydropyrazol-2-yl]-5H-thi-
azol-2-one (1) (0.01 mol) with the appropriate aromatic
aldehyde (0.01 mol) and sodium acetate (0.01 mol) in the
acetic acid medium were refluxed for 3-4 hours. After
cooling the reaction mixture formed precipitate was filtered
off and recrystallized.
5-Benzylidene-4-[3-(5-bromo-2-hydroxyphenyl)-5-phe-
nyl-3,4-dihydropyrazol-2-yl]thiazol-2-one (2a).
Yield: 72%; yellow solid; mp 242-244 °C, (DMF-EtOH
1:3). 1H NMR (400 MHz, CDCl3) δ 7.86 (d, 2Н, J 8.2 Hz,
Ar), 7.66 (m, 4Н, Ar), 7.40-7.55 (m, 5Н, Ar, СН=), 7.19 (d,
1Н, Ar), 7.04 (d, J 7.3 Hz, 1Н, Ar), 6.84 (d, J 8.0 Hz, 1Н,
Ar), 6.04 (dd, J 3.3, 20.0 Hz, 1Н, С5H-pyraz.), 3.98 (dd,
J 11.1, 20.0 Hz, 1Н, С4H-pyraz.), 3.26 (dd, J 3.3, 20.0 Hz,
1Н, С4H-pyraz.). Anal. Calcd. for C25H18BrN3O2S: C,
59.53; H, 3.60; N, 8.33. Found: C, 59.62; H, 3.68; N, 8.29.
4-[3-(5-Bromo-2-hydroxyphenyl)-5-phenyl-3,4-dihydro-
pyrazol-2-yl]-5-[(4-methoxyphenyl)methylene]thiazol-2-one
(2b).
Yield: 78%; yellow-orange solid; mp > 250 °C, (DMF-
EtOH 1:3). 1H NMR (400 MHz, CDCl3) δ 9.89 (s, 1Н, ОН),
7.88-7.93 (m, 2Н, Ar), 7.64 (d, J 8.0 Hz, 2Н, Ar), 7.50-7.54
(m, 3Н, Ar, СН=), 7.19 (d, 1Н, Ar), 7.06 (d, J 7.3 Hz, 3Н,
Ar), 6.83 (d, J 8.0 Hz, 1Н, Ar), 6.03 (dd, J 3.3, 20.0 Hz, 1Н,
С5H-pyraz.), 3.98 (dd, J 11.1, 20.0 Hz, 1Н, С4H-pyraz.),
3.29 (dd, J 5.0, 10.0 Hz, 1Н, С4H-pyraz.), 1.91 (s, 3Н,
СН3). Anal. Calcd. for C28H20BrN3O3S: C, 58.43; H, 3.77;
N, 7.86. Found: C, 58.50; H, 3.85; N, 7.80.
4-[3-(5-Bromo-2-hydroxyphenyl)-5-phenyl-3,4-dihydro-
pyrazol-2-yl]-5-[(3-nitrophenyl)methylene]thiazol-2-one
(2c).
Yield: 69%; yellow-orange solid; mp > 250 °C, (DMF-
EtOH 1:3). 1H NMR (400 MHz, CDCl3) δ 10.26 (s, 1Н,
ОН), 9.16 (s, 1Н, Ar), 8.36 (d, 2Н, J 8.0 Hz, Ar), 7.98-8.10
(m, 4Н, Ar), 7.60 (m, 3Н, Ar, СН=), 7.31 (d, 1Н, Ar), 7.20
(s, 1Н, Ar), 6.86 (d, J 8.0 Hz, 1Н, Ar), 6.02 (dd, J 3.3, 20.0
Hz, 1Н, С5H-pyraz.), 4.09 (dd, J 11.1, 20.0 Hz, 1Н, С4H-
pyraz.), 3.39 (dd, J 5.0, 10.0 Hz, 1Н, С4H-pyraz.). Anal.
A. P. Kryshchyshyn-Dylevych
47
Calcd. for C25H19BrN4O4S: C, 54.45; H, 3.47; N, 10.16.
Found: C, 54.60; H, 3.40; N, 10.03.
Biological tests
Anticancer in vitro screening methodology as well as
data interpretation are described in details at the NCI
Development Therapeutics Program site [20].
Cytotoxicity assays
The human hepatoma cell line (HepG2) was purchased
from the American Type Culture Collection (ATCC HB-
8065). The cells were cultured in Minimum Essential
Medium Eagle (MEME) (ATCC, USA). The murine
fibroblasts cell line (Balb/c 3T3 clone A31) (gift from
Department of Swine Diseases of the National Veterinary
Research Institute in Pulawy, Poland) was cultured in
Dulbecco`s Modified Eagle`s Medium (DMEM) (ATCC,
USA). The media were supplemented with 10% BCS
(Balb/c 3T3), 10% FBS (HepG2), 1% L-glutamine, 1%
antibiotic solution. The cells were maintained in 75 cm2 cell
culture flasks (NUNC) in a humidified incubator at 37 °C,
in an atmosphere of 5% CO2. The medium was refreshed
every two or three days and the cells were trypsinized by
0.25% trypsin-0.02% EDTA after reaching 70-80%
confluence. Single cell suspensions were prepared and
adjusted to a density of 2x105 cell/mL (HepG2) and 1x105
cell/mL for 24h, 48h exposition or 5x104 cell/mL for 72 h
exposition (Balb/c 3T3). The cell suspension was
transferred to 96-well plates (100 µl/well) and incubated for
24 h before the exposure to the studied compound. Stock
solution of the studied compound was prepared in DMSO
and diluted with culture medium to obtain a concentration
range from 10-5-102 µM. The cells were also exposed to the
reference drug – cisplatin used as internal laboratory
control. The final concentration of DMSO was 0.1% in the
medium and had no influence on cell growth. The medium
used for test solutions and in control preparation did not
contain serum and antibiotics. As negative control, cultured
cells were grown in the absence of study compound. Each
concentration was tested in six replicates with three
independent experiments. Cytotoxicity was assessed after
24, 48 and 72 h of exposure the cells to tested compound.
The medium was not changed during the incubation time.
MTT assay. The metabolic activity of living cells was
assessed by the measurement of the activity of
dehydrogenases [21].
NRU assay. The method is based on staining living cells
with neutral red which readily diffuses through the plasma
membrane and accumulates in lysosomes [22].
TCP assay. The assay was based upon staining total
cellular protein (proliferation) [23].
LDH leakage assay. The integrity of the plasma
membrane was assessed through the test of lactate
dehydrogenase (LDH) release [24], which was monitored
using the commercially available Cytotoxicity Detection Kit
(LDH) (Roche Diagnostics, Poland). The absorbance was
measured at microplate reader (Synergy HTXmulti-mode
reader (BioTek® Instruments Inc., USA)) at 570 nm, using
blank as a references. Cytotoxicity was expressed as a
percentage of the negative control (0.1% DMSO) [25].
Selectivity index.
To determine the cytotoxic selectivity of the tested
compound, the selectivity index (SI) was calculated
according to the following equation:
SI = CC50
no cancer cells/CC50
cancer cells
If a SI was found to be ≥ 3 the compound can be considered
to be selective [18].
A beneficial SI > 3.0 indicates a drug with efficacy
against tumor cells greater than toxicity against normal
cells. The results of the cytotoxicity assessment were
expressed as mean arithmetic values from three independent
experiments. The percentage of viability inhibition was
calculated in comparison with the untreated controls. The
CC20, CC50, CC80 values (cytotoxicity concentrations) are
the compound’s concentrations that inhibit the cell viability
by 20%, 50% and 80%were calculated by GraphPad Prism
5 software (San Diego, CA, USA) using nonlinear
regression.
Notes
Acknowledgements. I would like to thank National
Cancer Institute, Bethesda, MD, US for the anticancer
activity investigations and Department of Pharmacology
and Toxicology of the National Veterinary Research
Institute, Pulawy, Poland for the cytotoxicity studies. The
work was supported by the National Research Fund of
Ukraine (project № 2020.02/0035).
References
1. Rana, A.; Alex, J. M.; Chauhan, M.; Joshi, G.; Kumar, R. A review
on pharmacophoric designs of antiproliferative agents. Med. Chem.
Res. 2015, 24, 903-920.
2. Kaminskyy, D.; Kryshchyshyn, A.; Lesyk, R. 5-Ene-4-
thiazolidinones – An efficient tool in medicinal chemistry. Eur. J
Med. Chem. 2015, 140, 542-594.
3. Havrylyuk, D.; Roman, O.; Lesyk, R. Synthetic approaches, structure
activity relationship and biological applications for
pharmacologically attractive pyrazole/pyrazoline-thiazolidine-based
hybrids. Eur. J Med. Chem. 2016, 113, 145-166.
4. Zheng, W.; Degterev, A.; Hsu, E.; Yuan, J.; Yuan, C. Structure-
activity relationship study of a novel necroptosis inhibitor,
necrostatin-7. Bioorg. Med. Chem. Lett. 2008, 18, 4932-4935.
5. Isloor, A. M.; Sunil, D.; Shetty, P.; Malladi, S.; Pai, K. S. R.;
Maliyakki, N. Synthesis, characterization, anticancer, and antioxidant
activity of some new thiazolidin-4-ones in MCF-7 cells. Med. Chem.
Res. 2013, 22, 758-767.
6. Havrylyuk, D.; Zimenkovsky, B.; Vasylenko, O.; Zaprutko, L.;
Gzella, A.; Lesyk, R. Synthesis of novel thiazolone-based
compounds containing pyrazoline moiety and evaluation of their
anticancer activity. Eur. J Med. Chem. 2009, 44, 1396-1404.
7. Adjei, A. A.; Charron, M.; Rowinsky, E. K.; Svingen, P. A.; Miller,
J.; Reid, J. M.; Sebolt-Leopold, J.; Ames M. M.; Kaufmann, S. H.
Effect of pyrazoloacridine (NSC 366140) on DNA topoisomerases I
and II. Clin. Cancer Res. 1998, 4, 683-691.
8. Mascarenhas, J.; Hoffman, R. Ruxolitinib: the first FDA approved
therapy for the treatment of myelofibrosis. Clin. Cancer Res.
2012, 18, 3008-3014.
ISSN 1814-9758. Ukr. Bioorg. Acta, 2020, Vol. 15, N 2
48
9. Kryshchyshyn, A.; Kaminskyy, D.; Grellier, P.; Lesyk, R.
Thiazolidinone-Related Heterocyclic Compounds as Potential
Antitrypanosomal Agents. In Azoles-Synthesis, Properties,
Applications and Perspectives. IntechOpen, 2020.
10. Kryshchyshyn-Dylevych, A. P.; Zelisko, N. I.; Grellier, P.; Lesyk, R.
B. Preliminary evaluation of thiazolidinone- and pyrazoline-related
heterocyclic derivatives as potential antimalarial agents. Biopolym.
Cell. 2020, 36, 48-60.
11. Havrylyuk, D.; Zimenkovsky, B.; Karpenko, O.; Grellier, P.; Lesyk,
R. Synthesis of pyrazoline–thiazolidinone hybrids with trypanocidal
activity. Eur. J Med. Chem. 2014, 85, 245-254.
12. Havrylyuk, D.; Zimenkovsky, B.; Vasylenko, O.; Day, C. W.; Smee,
D. F.; Grellier, P.; Lesyk, R. Synthesis and biological activity
evaluation of 5-pyrazoline substituted 4-thiazolidinones. Eur. J Med.
Chem. 2013, 66, 228-237.
13. Kryshchyshyn, A.; Kaminskyy, D.; Karpenko, O.; Gzella, A.;
Grellier, P.; Lesyk, R. Thiazolidinone/thiazole based hybrids – New
class of antitrypanosomal agents. Eur. J Med. Chem. 2019, 174, 292-
308.
14. Bolognesi, M. L.; Cavalli, A. Multitarget drug discovery and
polypharmacology. Chem. Med. Chem. 2016, 11, 1190-1192.
15. Boyd, M. R.; Paull, K. D. Some practical considerations and
applications of the national cancer institute in vitro anticancer drug
discovery screen, Drug Dev. Res. 1995, 34, 91-109.
16. Boyd, M. R. in: Cancer Drug Discovery and Development, B.A
Teicher (Ed.), Humana Press, 1997, pp. 23-43.
17. Shoemaker, R. H. The NCI60 human tumour cell line anticancer drug
screen, Nat. Rev. Cancer. 2006, 6, 813-823.
18. Badisa, R. B.; Darling-Reed, S. F.; Joseph, P.; Cooperwood J. S.;
Latinwo L. M.; Goodman C. B. Selective cytotoxic activities of two
novel synthetic drugs on human breast carcinoma MCF-7 cells.
Anticancer Res. 2009, 29, 2993-2996.
19. Palaska, E.; Aytemir, M.; Uzbay, I. T.; Erol, D. Synthesis and
antidepressant activities of some 3, 5-diphenyl-2-pyrazolines. Eur. J
Med. Chem. 2001, 36, 539-543.
20. NCI-60 Human Tumor Cell Lines Screen. DTP Developmental
Therapeutics Program, NIH website [Internet]. Available from:
https://dtp.cancer.gov/discovery_development/nci-60/default.htm
(accessed on October 10, 2020).
21. Mosmann, T. Rapid colorimetric assay for cellular growth and
survival: application to proliferation and cytotoxicity assay. J.
Immunol. Methods. 1983, 65, 55-63.
22. Borenfreund, E.; Puerner, J. A. Toxicity determined in vitro by
morphological alterations and neutral red absorption. Toxicol. Lett.
1985, 24, 119-124.
23. Bradford,, M. M. A rapid and sensitive method for the quantitation of
microgram quantities of protein using the principle of dye binding.
Anal. Biochem. 1976, 72, 248-254.
24. Korzeniewski, C.; Calleawert, D.M. An enzyme-release assay for
natural cytotoxicity. J. Immunol. Methods. 1983, 64, 313-320.
25. Radko, L.; Stypuła-Trębas, S.; Posyniak, A.; Żyro D.; Ochocki, J.
Silver(I) Complexes of the pharmaceutical agents metronidazole and
4-hydroxymethylpyridine: comparison of cytotoxic profile for
potential clinical application. Molecules. 2019, 24, 1949.
Деякі фармакологічні властивості
4-[3-(5-бромо-2-гідроксифеніл)-5-феніл-3,4-дигідропіразол-2-іл]-5Н-тіазол-2-ону
А. П. Крищишин-Дилевич*
Львівський національний медичний університет імені Данила Галицького, вул. Пекарська, 69, Львів, 79010, Україна
Резюме: Розроблено та синтезовано ряд піразоліл-тіазолідинонів як потенційних біологічно активних сполук. Дослідження протипухлинної
активності 4-[3-(5-бром-2-гідроксифеніл)-5-феніл-3,4-дигідропіразол-2-іл]-5Н-тіазол-2-ону (1) виявило його високі антипроліферативні
властивості щодо панелі більш, ніж 50-ти ракових клітинних ліній з найнижчою концентрацією пригнічення росту (GI50) обчисленою для лінії
лейкемії SR (0,0351 мкМ) та лінії раку яйників OVCAR-3 (0,248 мкМ). Крім того, піразоліл-тіазолідинон 1 повністю пригнічував ріст (TGI) ліній
лейкемії CCRF-CEM, HL-60, MOLT-4 та SR при субмікромолярних значеннях концентрації. Крім ліній лейкемії, чутливими до дії сполуки 1 були
лінії недрібноклітинного раку легень NCI-H322M та EKVX, лінія раку товстого кишківника HCT-116, раку яйників OVCAR-3 та раку молочної
залози MCF7. Також було встановлено, що піразоліл-тіазолідинон 1 інгібує ріст Trypanosoma brucei brucei на 98,8% у концентрації 10 мкг/мл, що
свідчить про перспективність розробки даного агенту в рамках концепції «поліфармакологічних лікарських засобів». Поглиблене д ослідження
цитотоксичності сполуки 1 на клітинах гепатоцелюлярної карциноми людини HepG2 та нормальних мишачих фібробластах Balb/c3T3 у MTT,
NRU, TPC та LDH тестах показало, що нормальні клітини менш чутливі до дії піразоліл-тіазолідинону 1, ніж ракові. Відповідно до значень CC50,
першим етапом протипухлинної дії сполуки 1 було пошкодження клітинних мембран, що призводить до інгібування мітохондріальної
активності, з послідовним інгібуванням проліферації та лізосомальної активності ракових клітин. Для 4-[3-(5-бром-2-гідроксифеніл)-5-феніл-3,4-
дигідропіразол-2-іл]-5Н-тіазол-2-ону (1) обчислено індекси селективності, що були вищими у тесті LDH, ніж такі для препарату порівняння
цисплатину.
Ключові слова: піразолін-тіазолідинонові гібриди; протипухлинна активність; антитрипаносомна активність; цитотоксичність.
Notes
Acknowledgements. I would like to thank National Cancer Institute, Bethesda, MD, US for the anticancer activity investigations and Department of Pharmacology and Toxicology of the National Veterinary Research Institute, Pulawy, Poland for the cytotoxi...
|
| id | oai:ojs2.bioorganica.com.ua:article-40 |
| institution | Ukrainica Bioorganica Acta |
| keywords_txt_mv | keywords |
| language | English |
| last_indexed | 2026-07-20T01:00:55Z |
| publishDate | 2020 |
| publisher | V.P. Kukhar Institute of Bioorganic Chemistry and Petrochemistry of the National Academy of Sciences of Ukraine |
| record_format | ojs |
| resource_txt_mv | bioorganicacomua/2f/278f05325bd8a4795284cd8022307f2f.pdf |
| spelling | oai:ojs2.bioorganica.com.ua:article-402026-07-19T14:56:53Z Some pharmacological properties of 4-[3-(5-bromo-2-hydroxyphenyl)-5-phenyl-3,4-dihydropyrazol-2-yl]-5H-thiazol-2-one Деякі фармакологічні властивості 4-[3-(5-бромо-2-гідроксифеніл)-5-феніл-3,4-дигідропіразол-2-іл]-5Н-тіазол-2-ону Kryshchyshyn-Dylevych, Anna pyrazolyl thiazolinone hybrids antitumor activity antitrypanosomal activity cytotoxicity піразолін-тіазолідинонові гібриди протипухлинна активність антитрипаносомна активність цитотоксичність A series of 3,5-diaryl pyrazolyl thiazolinones were designed and synthesized as potential biologically active compounds. The study of anticancer activity of 4-[3-(5-bromo-2-hydroxyphenyl)-5-phenyl-3,4-dihydropyrazol-2-yl]-5H-thiazol-2-one (1) revealed its high antiproliferative activity against a panel of cancer cells with the lowest growth inhibition concentration (GI50) towards leukemic cell line SR (0.0351 µМ) and ovarian cancer cell line OVCAR-3 (0.248 µМ). It was also found that pyrazolyl thiazolinone 1 inhibited growth of Trypanosoma brucei brucei by 98,8% at a concentration of 10 µg/mL. The in-depth cytotoxicity study of compound 1 on human hepatocellular carcinoma HepG2 cells and non-tumorigenic murine fibroblast Balb/c 3T3 in MTT, NRU, TPC and LDH assays showed that normal cells were less sensitive to compound 1 than the cancer cells; its action had led to a disintegration of the cell membrane, inhibition of mitochondrial and lysosomal activity, and proliferation of cancer cells. The highest selectivity were detected in the LDH assay Розроблено та синтезовано ряд піразоліл-тіазолідинонів як потенційних біологічно активних сполук. Дослідження протипухлинної активності 4-[3-(5-бром-2-гідроксифеніл)-5-феніл-3,4-дигідропіразол-2-іл]-5Н-тіазол-2-ону (1) виявило його високі антипроліферативні властивості щодо панелі більш, ніж 50-ти ракових клітинних ліній з найнижчою концентрацією пригнічення росту (GI50) обчисленою для лінії лейкемії SR (0,0351 мкМ) та лінії раку яйників OVCAR-3 (0,248 мкМ). Крім того, піразоліл-тіазолідинон 1 повністю пригнічував ріст (TGI) ліній лейкемії CCRF-CEM, HL-60, MOLT-4 та SR при субмікромолярних значеннях концентрації. Крім ліній лейкемії, чутливими до дії сполуки 1 були лінії недрібноклітинного раку легень NCI-H322M та EKVX, лінія раку товстого кишківника HCT-116, раку яйників OVCAR-3 та раку молочної залози MCF7. Також було встановлено, що піразоліл-тіазолідинон 1 інгібує ріст Trypanosoma brucei brucei на 98,8% у концентрації 10 мкг/мл, що свідчить про перспективність розробки даного агенту в рамках концепції «поліфармакологічних лікарських засобів». Поглиблене дослідження цитотоксичності сполуки 1 на клітинах гепатоцелюлярної карциноми людини HepG2 та нормальних мишачих фібробластах Balb/c3T3 у MTT, NRU, TPC та LDH тестах показало, що нормальні клітини менш чутливі до дії піразоліл-тіазолідинону 1, ніж ракові. Відповідно до значень CC50, першим етапом протипухлинної дії сполуки 1 було пошкодження клітинних мембран, що призводить до інгібування мітохондріальної активності, з послідовним інгібуванням проліферації та лізосомальної активності ракових клітин. Для 4-[3-(5-бром-2-гідроксифеніл)-5-феніл-3,4-дигідропіразол-2-іл]-5Н-тіазол-2-ону (1) обчислено індекси селективності, що були вищими у тесті LDH, ніж такі для препарату порівняння цисплатину V.P. Kukhar Institute of Bioorganic Chemistry and Petrochemistry of the National Academy of Sciences of Ukraine 2020-12-30 Article Article application/pdf https://bioorganica.com.ua/index.php/journal/article/view/40 10.15407/bioorganica2020.02.041 Ukrainica Bioorganica Acta; Vol. 15 No. 2 (2020): Ukrainica Bioorganica Acta; 41-48 Ukrainica Bioorganica Acta; Том 15 № 2 (2020): Ukrainica Bioorganica Acta; 41-48 1814-9766 1814-9758 10.15407/bioorganica2020.02 en https://bioorganica.com.ua/index.php/journal/article/view/40/39 Copyright (c) 2020 Anna Kryshchyshyn-Dylevych https://creativecommons.org/licenses/by/4.0 |
| spellingShingle | піразолін-тіазолідинонові гібриди протипухлинна активність антитрипаносомна активність цитотоксичність Kryshchyshyn-Dylevych, Anna Деякі фармакологічні властивості 4-[3-(5-бромо-2-гідроксифеніл)-5-феніл-3,4-дигідропіразол-2-іл]-5Н-тіазол-2-ону |
| title | Деякі фармакологічні властивості 4-[3-(5-бромо-2-гідроксифеніл)-5-феніл-3,4-дигідропіразол-2-іл]-5Н-тіазол-2-ону |
| title_alt | Some pharmacological properties of 4-[3-(5-bromo-2-hydroxyphenyl)-5-phenyl-3,4-dihydropyrazol-2-yl]-5H-thiazol-2-one |
| title_full | Деякі фармакологічні властивості 4-[3-(5-бромо-2-гідроксифеніл)-5-феніл-3,4-дигідропіразол-2-іл]-5Н-тіазол-2-ону |
| title_fullStr | Деякі фармакологічні властивості 4-[3-(5-бромо-2-гідроксифеніл)-5-феніл-3,4-дигідропіразол-2-іл]-5Н-тіазол-2-ону |
| title_full_unstemmed | Деякі фармакологічні властивості 4-[3-(5-бромо-2-гідроксифеніл)-5-феніл-3,4-дигідропіразол-2-іл]-5Н-тіазол-2-ону |
| title_short | Деякі фармакологічні властивості 4-[3-(5-бромо-2-гідроксифеніл)-5-феніл-3,4-дигідропіразол-2-іл]-5Н-тіазол-2-ону |
| title_sort | деякі фармакологічні властивості 4-[3-(5-бромо-2-гідроксифеніл)-5-феніл-3,4-дигідропіразол-2-іл]-5н-тіазол-2-ону |
| topic | піразолін-тіазолідинонові гібриди протипухлинна активність антитрипаносомна активність цитотоксичність |
| topic_facet | pyrazolyl thiazolinone hybrids antitumor activity antitrypanosomal activity cytotoxicity піразолін-тіазолідинонові гібриди протипухлинна активність антитрипаносомна активність цитотоксичність |
| url | https://bioorganica.com.ua/index.php/journal/article/view/40 |
| work_keys_str_mv | AT kryshchyshyndylevychanna somepharmacologicalpropertiesof435bromo2hydroxyphenyl5phenyl34dihydropyrazol2yl5hthiazol2one AT kryshchyshyndylevychanna deâkífarmakologíčnívlastivostí435bromo2gídroksifeníl5feníl34digídropírazol2íl5ntíazol2onu |