Mendeleev Commun., 2017, 27, 228–230
was required in comparison with the earlier established conditions.
C(32)
A gentle reagent was found for the reduction of nitro group in
isoxazole core. Two biotests revealed moderate antimitotic activity
of compounds 1a and 1e to human lung carcinoma A549 cell
line. These compounds are of interest for additional investigation
of the mechanism of their antiproliferative properties.
O(13)
O(31)
C(4)
C(3)
C(5)
C(2)
C(11)
C(1)
C(6)
C(13)
O(41)
O(12)
C(17)
C(16)
C(18)
N(14)
O(51)
C(42)
C(52)
This work was supported by the Russian Foundation for
Basic Research (project nos. 14-03-00469 and 15-03-04894),
the Russian Academy of Sciences (programme nos. 8P and 9)
and the German Academic Exchange Service under the auspices
of a collaborative agreement between Moscow and Rostock
Universities. This study was fulfilled using a STOE STADI VARI
PILATUS-100K diffractometer purchased by MSU Develop-
ment Program.
O(15)
N(16)
Figure 2 The general view of 3,4,5-trimethoxybenzyl 5-aminoisoxazole-
3-carboxylate 1a in representation of atoms by thermal ellipsoids (p = 50%).
Novel substituted 5-aminoisoxazoles were examined for
their cytotoxicity to epithelial lung carcinoma cancer cells
A549 in a standard colorimetric MTT assay using the procedure
described17–19 (Table 2; for details, see Online Supplementary
Materials). Their cytotoxicity turned out to be inferior to that of
podophyllotoxin or A-105972. Nevertheless, two compounds
of the series, namely 1a and 1e, were moderately cytotoxic with
EC50 values in the micromolar concentration range. The data
obtained demonstrated that the introduction of methyl substituent
either to the isoxazole or to the linker chain (compounds 1b–d,f)
led to a decrease in cytotoxicity. Interestingly, the equal EC50
values for 1a and 1e indicated the equivalence of the trimethoxy-
phenyl and methylenedioxyphenyl rings for the cytotoxic activity
of these compounds.
Online Supplementary Materials
Supplementary data associated with this article can be found
in the online version at doi: 10.1016/j.mencom.2017.05.003.
References
1 G. A. Patani and E. J. LaVoie, Chem. Rev., 1996, 96, 3147.
2 N. A. Meanwell, J. Med. Chem., 2011, 54, 2529.
3 S. G. Zlotin, A. M. Churakov, O. A. Luk’yanov, N. N. Makhova, A. Yu.
Sukhorukov and V. A. Tartakovsky, Mendeleev Commun., 2015, 25, 399.
4 Y.A. Volkova, E. B.Averina,Y. K. Grishin, P. Bruheim, T. S. Kuznetsova
and N. S. Zefirov, J. Org. Chem., 2010, 75, 3047.
We also studied the effect of compounds 1a, 1e and 1f
(100 mm) on cell growth using direct cell counting by microscopy
over 24, 48 and 72 h of culturing (Figure 3). The compounds
strongly inhibited cell proliferation, and the extent of inhibition
correlated with the cytotoxicity of compounds 1a,e,f (see Table 2).
Thus, cell growth experiments confirmed the MTT data.
In conclusion, a three-step synthetic protocol was accom-
plished for the preparation of substituted 5-aminoisoxazoles
with polymethoxyphenyl- and methylenedioxyphenyl moieties.
The optimization of the first and the third steps of the protocol
5 E. B. Averina, Y. V. Samoilichenko, Y. A. Volkova, Y. K. Grishin, V. B.
Rybakov, A. G. Kutateladze, M. E. Elyashberg, T. S. Kuznetsova and
N. S. Zefirov, Tetrahedron Lett., 2012, 53, 1472.
6 E. B. Averina, D. A. Vasilenko,Y. V. Samoilichenko,Y. K. Grishin, V. B.
Rybakov, T. S. Kuznetsova and N. S. Zefirov, Synthesis, 2014, 46, 1107.
7 T. L. Nguyen, C. McGrath,A. R. Hermone, J. C. Burnett, D. W. Zaharevitz,
B. W. Day, P. Wipf, E. Hamel and R. Gussio, J. Med. Chem., 2005, 48,
6107.
8 O. N. Zefirova, A. G. Diikov, N. V. Zyk and N. S. Zefirov, Russ. Chem.
Bull., Int. Ed., 2007, 56, 680 (Izv. Akad. Nauk, Ser. Khim., 2007, 655).
9 J. R. Wu-Wong, J. D. Alder, L. Alder, D. J. Burns, E. K.-H. Han, B. Credo,
S. K. Tahir, B. D. Dayton, P. J. Ewing and W. J. Chiou, Cancer Res.,
2001, 61, 1486.
10 W. Wang, Y. Li, M. Sun, C. Zhou, Y. Zhang, Y. Li and Q. Yang, Chem.
Commun., 2012, 48, 6040.
11 E. B. Averina, D. A. Vasilenko, Y. A. Gracheva, Y. K. Grishin, E. V.
Radchenko, V. V. Burmistrov, G. M. Butov, M. E. Neganova, T. P. Serkova,
O. M. Redkozubova, E. F. Shevtsova, E. R. Milaeva, T. S. Kuznetsova
and N. S. Zefirov, Bioorg. Med. Chem., 2016, 24, 712.
12 J. S.Yadav, G. S. Reddy, D. Srinivas and K. Himabindu, Synth. Commun.,
1998, 28, 2337.
13 M. Bruder, D. B. Vendramini-Costa, J. E. de Carvalho and R. A. Pilli,
Bioorg. Med. Chem., 2013, 21, 5107.
Table 2 Cytotoxicity of compounds 1a–f to human lung carcinoma cell line
A549.
Compound
Cytotoxicity (EC50, mm) Cell growth inhibition (%)a
1a
1b
1c
1d
1e
1f
38±4
>100
>100
>100
35±5
93±7
91
–
–
–
100
55
–
14 V. K. Yadav and K. Ganesh Babu, J. Org. Chem., 2004, 69, 577.
15 A. Pascual, Helv. Chim. Acta, 1991, 74, 531.
16 M. Weiwer, J. A. Bittker, T. A. Lewis, K. Shimada, W. S. Yang, L. Mac-
Pherson, S. Dandapani, M. Palmer, B. R. Stockwell, S. L. Schreiber and
B. Munoz, Bioorg. Med. Chem. Lett., 2012, 22, 1822.
17 O. N. Zefirova, H. Lemcke, M. Lantow, E. V. Nurieva, B. Wobith, G. E.
Onishchenko, A. Hoenen, G. Griffiths, N. S. Zefirov and S. A. Kuznetsov,
ChemBioChem, 2013, 14, 1444.
Podophyllotoxin 0.04
aAfter 48 h of culturing.
300
250
200
150
100
50
DMSO
1f
18 A. A. Beloglazkina, B. Wobith, E. S. Barskaia, N.A. Zefirov,A. G. Majouga
,
1e
1a
E. K. Beloglazkina, N. V. Zyk, S. A. Kuznetsov and O. N. Zefirova, Med.
Chem. Res., 2016, 25, 1239.
19 O. N. Zefirova, E. V. Nurieva, H. Lemcke, A. A. Ivanov, N. V. Zyk,
D. G. Weiss, S. A. Kuznetsov and N. S. Zefirov, Mendeleev Commun.,
2008, 18, 183.
0
0
24
48
72
t/h
Figure 3 Effect of compounds 1a, 1e and 1f on cell growth in culture. The
cells were incubated with 100 mm of each compound and after resuspension
were counted by phase-contract microscopy using hemocytometer. 1% DMSO
was used as a control.
Received: 2nd September 2016; Com. 16/5037
– 230 –