Synthesis of new compounds in the series of aryl-substituted ureas with cytotoxic and antioxidant activity

Article abstract of DOI:10.1016/j.mencom.2020.03.007

A series of aryl-substituted ureas and carbamates containing chlorinated aromatic and modified imidazolidinone moieties were synthesized. These compounds were found to be cytotoxic to breast cancer cell line MDA-MB-231, glioblastoma U-87 MG and neuroblast

Full text of DOI:10.1016/j.mencom.2020.03.007

Mendeleev
Communications
Mendeleev Commun., 2020, 30, 153–155
Synthesis of new compounds in the series of aryl-substituted ureas
with cytotoxic and antioxidant activity
Antonida V. Kalistratova,a Leonid V. Kovalenko,a Maxim S. Oshchepkov,*a Alina M. Gamisoniya,b
Tatiana S. Gerasimova,a Yuri A. Demidova and Mikhail G. Akimovb
a D. I. Mendeleev University of Chemical Technology of Russia, 125047 Moscow, Russian Federation.
E-mail: maxim.os@mail.ru
b M. M. Shemyakin–Yu. A. Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences,
117997 Moscow, Russian Federation. E-mail: akimovmike@yandex.ru
DOI: 10.1016/j.mencom.2020.03.007
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A series of aryl-substituted ureas and carbamates containing
chlorinated aromatic and modified imidazolidinone moieties
were synthesized. These compounds were found to be
cytotoxic to breast cancer cell line MDA-MB-231,
glioblastoma U-87 MG and neuroblastoma SH-SY5Y, but
not to melanoma A-375.
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Keywords: synthetic cytokinins, substituted ureas, anti-stress effect, cytotoxicity, oxidative stress.
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The search for anticancer drugs among existing cytokinins and
cytokinin-like compounds is currently of considerable interest.1
Cytokinins are small molecule biologically active compounds of
phytohormone group that play an important role at all stages of
plant growth and development.2,3 In addition to the action exerted
on plants, the biological activity of natural and synthetic
cytokinins towards animals was reported.4,5 The key aspect of
this activity is a cytotoxic effect,6 which is being studied on a
number of malignant cells lines. Another interesting activity of
cytokinin-like compounds is the ability to act as antioxidants,
which can be of use in cosmetology.7
H
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Cl
Kartolin-2
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EDU analogue
The antitumor activity of cytokinins is associated with
regulation of cell proliferation and differentiation, similar to that
occurring in plants. Thus, both N6-substituted adenines and
cytokinin-like arylureas showed positive in vitro and in vivo
results against glioblastoma,8,9 rhabdomyosarcoma, breast
cancer, CNS tumors, colon cancer, lung cancer, leukemia,
melanoma, prostate, ovaries and kidney cancers.6,10,11 The
mechanism of cytokinin action on tumor cells is thought to be
based on stopping cell division, mainly by blocking various
cyclin-dependent kinases,12 inducing genotoxic stress in
oncogenes, and activating the N-terminal kinase c-Jun.2
Figure 1 Structure of cytokinin-like compounds.
We suggested that the modification of EDU structure by
introducing a chlorine atom into the aromatic ring may be of
interest, because it reproduces a moiety similar to kartolin-2.
Imidazolidinone 1 was obtained by the condensation of
diethylene triamine with urea in 65% yield. Compound 2 is
commercially available as a 75% aqueous solution. It was
concentrated and dried by the azeotropic distillation of water
with CCl4 prior to synthesis.
Compounds 3–7 were produced by reaction of key compounds
1 and 2 with corresponding arylisocyanates in the presence of
triethylamine in anhydrous toluene (for arylureas) or in
acetonitrile (for arylcarbamates) in yields ranging from 35 to
68% (Scheme 1; for details, see Online Supplementary
Materials).
The resulting compounds were tested for cytotoxicity
against four human tumor cell lines: melanoma A-375,
glioblastoma U-87 MG, breast cancer MDA-MB-231 and
neuroblastoma SH-SY5Y. Cells were incubated with the
compounds for 24 h, and the cell viability was determined by
the MTT assay.
Arylureas like kartolin-2 and structurally related compound,
N-[2-2-oxoimidazolidin-1-yl)ethyl]-N'-phenylurea
(EDU
analogue) (Figure 1) are similar in biological properties to
natural adenine-type cytokinins, but are more synthetically
available. Both of these compounds exhibit cytokinin-like
activity; the ability of the latter to protect plants from the action
of ozone by slowing down defoliation was also noted.13,14
Similar antioxidant effects are observed in animal cells.15
Here we report the synthesis of aryl-substituted ureas and
carbamates structurally close to EDU and kartolin-2 and the
study of their biological activity.
© 2020 Mendeleev Communications. Published by ELSEVIER B.V.
on behalf of the N. D. Zelinsky Institute of Organic Chemistry of the
Russian Academy of Sciences.
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Mendeleev Commun., 2020, 30, 153–155
fragment or its analogue and this is consistent with the published
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cytotoxicity data.16,17 Regardless of low cytotoxicity, a selectivity
of action was present, which makes further search for analogous
compounds with increased activity promising.
i or ii
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2 X = O
Since there are indications of a possible antioxidant effect of
cytokinin analogues in literature, the protective effect of
synthesized compounds was studied in three models of oxidative
stress caused by reactive oxygen species. These models were
protection against H2O2, chemical hypoxia induced by CoCl2,18
and low glucose conditions19 (Figure 3). The toxic agent was
added simultaneously with the tested compounds and incubated
with the cells for 24 h, after that cell viability was determined
using the MTT assay. Previously developed dose-response
curves for H2O2 and CoCl2 provided a basis on which the definite
concentrations were selected for causing the death of 30–40% of
cells to avoid nonspecific toxicity.20
The study of the protective effect showed that a significant
increase in cell viability was observed only under oxidative
stress conditions for the compound containing chlorine only in
para-position (4). On the other hand, compound 5 with the
same modification did not have a protective effect. The
presence of such neuroprotective activity is consistent with
previously reported data for halogen-containing 4-(cycloalkyl)
piperidines.21
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N
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3–7
R1 R2
Yield (%)
35
X
NH
O
NH
O
H
H
H
Cl
H
3
Cl 54
Cl 53
Cl 39
Cl 68
4
5
6
7
NH Cl
Scheme 1 Reagents and conditions: i, Et3N, MeCN, room temperature,
24 h (4, 6); ii, Et3N, toluene, 5 °C, 24 h (3, 5, 7).
It was found that several compounds were cytotoxic for breast
cancer and neuroblastoma cell lines (Figure 2), decreasing
viability to 50–60% at 100 μm. Only parent compound 3 was
cytotoxic for U-87 MG line, causing 40% viability decrease at
100 μm. We can conclude that introduction of chlorine into the
benzene ring significantly reduced the cytotoxicity for
glioblastoma. Beside that, the decrease of substituent size
(replacing imidazolidinone with ethanol) and the introduction of
chlorine into para-position of the benzene ring led to a loss of
the activity.
Thus, it has been found that the introduction of chlorine into
the aromatic ring of cytokinin analogues significantly reduces
cytotoxicity, however gives the molecule the capability of
protecting cells from oxidative stress induced by H2O2.
This work was supported by the Russian Foundation for Basic
Research (grant no. 19-03-00492 A).
Online Supplementary Materials
Leaders in the cytotoxicity tests were compounds 3 and 7.
Their distinguishing feature is the presence of an imidazolidinone
Supplementary data associated with this article can be found
in the online version at doi: 10.1016/j.mencom.2020.03.007.
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Figure 2 Cytotoxic effect of the synthesized compounds 3–7 against different lines of the tumor cells (a) A-375, (b) MDA-MB-231, (c) U-87 MG and
(d) SH-SY5Y. Incubation time 24 h, MTT assay, mean SD (N = 3).
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Mendeleev Commun., 2020, 30, 153–155
References
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1 C. Laezza, M. G. Caruso, T. Gentile, M. Notarnicola, A. M. Malfitano,
T. Di Matola, C. Messa, P. Gazzerro and M. Bifulco, Int. J. Cancer,
2009, 124, 1322.
2 L. V. Kovalenko, A. V. Kalistratova, M. S. Oshchepkov,
E. A. Glukhoedova, I. N. Soloviova, M. M. Vorobiov, K. A. Kochetkov
and S. D. Karakotov, Butlerovskie Soobshcheniya, 2017, 50, 146
(in Russian).
ꢁ0
0
3 D. Zalabák, H. Pospíšilová, M. Šmehilová, K. Mrízová, I. Frébort and
P. Galuszka, Biotechnol. Adv., 2013, 31, 97.
4 E. M. Savelieva, V. E. Oslovsky, D. S. Karlov, N. N. Kurochkin,
I. A. Getman, S. N. Lomin, G. V. Sidorov, S. N. Mikhailov,
D. I. Osolodkin and G. A. Romanov, Phytochemistry, 2018, 149, 161.
5 M. S. Oshchepkov, A. V. Kalistratova, E. M. Savel’eva, G. A. Romanov,
N. A. Bystrova and K. A. Kochetkov, Russ. Chem. Rev., 2020, 89,
doi: 10.1070/rcr4921.
6 J. Voller, M. Zatloukal, R. Lenobel, K. Dolezal, T. Béres, V. Krystof,
L. Spíchal, P. Niemann, P. Dzubák, M. Hajdúch and M. Strnad,
Phytochemistry, 2010, 71, 1350.
ConcentrationꢈµM
ꢀ
ꢂ
ꢃ
ꢁ
ꢄ
ꢇ ꢇ ꢇ
ꢀ00
7 E. M. Othman, M. Naseem, E. Awad and T. Dandekar and H. Stopper,
PLoS ONE, 2016, 11, e0168386.
8 E. Ciaglia, M. Abate, C. Laezza, S. Pisanti, M. Vitale, V. Seneca,
G. Torelli, S. Franceschelli, G. Catapano, P. Gazzerro and M. Bifulco,
Int. J. Cancer, 2017, 140, 959.
9 E. Ciaglia, C. Laezza, M. Abate, S. Pisanti, R. Ranieri, A. D’Alessandro,
P. Picardi, P. Gazzerro and M. Bifulco, Int. J. Cancer, 2018, 142, 176.
10 J. Voller, T. Béres, M. Zatloukal, P. A. Kaminski, P. Niemann, K. Doležal,
P. Džubák, M. Hajdúch and M. Strnad, Phytochemistry, 2017, 136, 156.
11 J. Voller, M. Zatloukal, R. Lenobel, K. Dolezal, T. Béres, V. Krystof,
L. Spíchal, P. Niemann, P. Dzubák, M. Hajdúch and M. Strnad,
Phytochemistry, 2010, 71, 1350.
12 P. Kittakoop, Stud. Nat. Prod. Chem., 2015, 44, 251.
13 E. H. Lee and C. M. Chen, Physiol. Plant., 1982, 56, 486.
14 S. Singh and S. B. Agrawal, Water, Air, Soil Pollut., 2011, 217, 283.
15 J. S. Kerr and G. A. Boswell, US Patent 5001141A, 1991.
16 C. Anselmi, A. Ettorre, M. Andreassi, M. Centini, P. Neri and
A. Di Stefano, Biochem. Pharmacol., 2002, 63, 437.
ꢁ0
0
ConcentrationꢈµM
ꢀ
ꢂ
ꢃ
ꢁ
ꢄ
ꢀ00
17 D. G. Spindola,A. Hinsberger,V. M. deSouzaAntunes, L. F. G. Michelin,
C. Bincoletto and C. R. Oliveira, Braz. J. Pharm. Sci., 2018, 54, 1.
18 Y. Wang, Z. Tang, R. Xue, G. K. Singh, W. Liu, Y. Lv and L. Yang,
Mol. Cell. Biochem., 2012, 360, 235.
ꢁ0
0
19 N. Jelluma, X. Yang, D. Stokoe, G. I. Evan, T. B. Dansen and
D. A. Haas-Kogan, Mol. Cancer Res., 2006, 4, 319.
20 M. G. Akimov, A. M. Ashba, E. V. Fomina-Ageeva, N. M. Gretskaya,
N. F. Myasoedov and V. V. Bezuglov, Dokl. Biochem. Biophys., 2019,
485, 141 (Dokl. Akad. Nauk, 2019, 485, 625).
ConcentrationꢈµM
21 P. Lardenois, J. Frost, P. Pasau, P. George, M. C. Renones, R. Bartsch,
W.-T. Li, P. Magat and R. Dupont, US Patent WO 009309, 1997.
Figure 3 Protective effect of synthesized compounds 3–7 in stress models.
(a) Chemical hypoxia (800 μm CoCl2), (b) oxidative stress (1200 μm H2O2),
(c) hypoglycemia (1 mm glucose concentration in cultural medium).
Incubation time 24 h, MTT assay, mean SD (N = 3); * – statistically
significant difference from control, p < 0.05, ANOVA with Dunnett’s post-
test.
Received: 9th September 2019; Com. 19/6027
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