Benzo[e][1,2,4]triazino[2,3-c][1,2,3]triazin-2-ones electro-deficient heterocyclic compounds with promising anticancer activity

Article abstract of DOI:10.1515/hc-2015-0190

The synthesis and antitumor activity of substituted benzo[e][1,2,4]triazino[2,3-c][1,2,3]triazin-2-ones-novel electro-deficient tricyclic compounds-are described. These compounds were prepared by treatment of 3-(2-amino-3-R²-5-R³-phenyl)-6-R¹-1,2,4-triazin-5(2H)-ones with sodium nitrite in acetic acid. Spectral properties of synthesized compounds were studied and compared with spectral data of known 3-R-8-R¹-10-R²-2H-[1,2,4]triazino[2,3-c]quinazolin-2-ones. These compounds are promising antitumor agents. The most active anticancer compound 3d was studied in dose-depended anticancer activity assay and its selective growth inhibition against breast cancer MDA-MB-468 cell line was established (GI₅₀=0.41 μm).

Full text of DOI:10.1515/hc-2015-0190

Heterocycl. Commun. 2016; 22(3): 137–141
Olexii Y. Voskoboynik*, Sergiy I. Kovalenko and Svetlana V. Shishkina
Benzo[e][1,2,4]triazino[2,3-c][1,2,3]triazin-2-ones
electro-deficient heterocyclic compounds with
promising anticancer activity
DOI 10.1515/hc-2015-0190
Received September 2, 2015; accepted April 26, 2016; previously
published online May 27, 2016
agents [12]. Considering our experience in the search for
antimicrobial, fungicidal and anticancer agents among
[1,2,4]triazino[2,3-c]quinazolines [13–16] and the reported
efficiency of bioisosteric replacement strategies [17, 18] it
was decided to elaborate synthetic protocol for series of
3-R1-8-R2-10-R3-2H-benzo[e][1,2,4]triazino[2,3-c][1,2,3]tria-
zin-2-ones. These compounds may be regarded as isosteric
analogs of previously described heterocyclic compounds
with promising antitumor activity [13–16].
Abstract: The synthesis and antitumor activity of sub-
stituted benzo[e][1,2,4]triazino[2,3-c][1,2,3]triazin-2-ones
– novel electro-deficient tricyclic compounds – are
described. These compounds were prepared by treatment
of
3-(2-amino-3-R2-5-R3-phenyl)-6-R1-1,2,4-triazin-5(2H)-
ones with sodium nitrite in acetic acid. Spectral properties
of synthesized compounds were studied and compared
with spectral data of known 3-R-8-R1-10-R2-2H-[1,2,4]
triazino[2,3-c]quinazolin-2-ones. These compounds are Results and discussion
promising antitumor agents. The most active anticancer
compound 3d was studied in dose-depended anticancer
Chemistry
activity assay and its selective growth inhibition against
breast cancer MDA-MB-468 cell line was established
(GI50ꢀ=ꢀ0.41 μm).
Keywords: anticancer activity; benzo[e][1,2,4]triazino[2,3- for synthesis of target 3-R1-8-R2-10-R3-2H-benzo[e][1,2,4]
c][1,2,3]triazines; heterocyclic compounds.
Substituted
3-(2-amino-3-R2-5-R3-phenyl)-6-R1-1,2,4-tri-
azin-5(2H)-ones 2a–j were used as starting materials
triazino[2,3-c][1,2,3]triazin-2-ones (3a–j, Scheme 1). The
starting anilines were obtained according to a known
synthetic protocol [19] via transformation of 3-R-8-R1-10-R2-
2H-[1,2,4]triazino[2,3-c]quinazolin-2-ones 1a–j [20].
Purity of the synthesized compounds was analyzed
by LC-MS (APCI) method and their structure was estab-
Introduction
Derivatives of 1,2,4- and 1,3,5-triazines show anticancer,
antibacterial and antifungal activity [1–6]. However, few
papers are devoted to the chemistry and evaluation of
biological activity of 1,2,3-triazines and their condensed
derivatives. Oxidation of N-aminopyrazoles is used to
synthesize 1,2,3-triazines [7, 8] and treatment of aromatic
o–aminonitriles and o-aminohydrazides with nitrous acid
or butyl nitrite yields condensed derivatives [9–11]. This
chemistry was used for the synthesis of novel antifungal
1
lished by using physicochemical methods including H
13
NMR, C NMR, IR and mass spectrometry. In particular,
the protons in positions 8 and 10 of 3a–j are significantly
deshielded in comparison to the corresponding protons
of [1,2,4]triazino[2,3-c]quinazoline system [20]. Signals
caused by the presence of a substituent in position 3 are
also registered in proper fields. The carbons are also more
deshielded in comparison to those of previously described
[1,2,4]triazino[2,3-c]quinazolines [20]. The mass spectrum
of compound 3b is characterized by a molecular ion peak
of low intensity (2%). One of the mass fragmentation
routes includes elimination of molecular nitrogen (m/z
247) from the molecular ion.
*Corresponding author: Olexii Y. Voskoboynik, Zaporizhia State
Medical University, Mayakovsky Ave., 26, Zaporozhia 69035,
Ukraine, e-mail: a.yu.voskoboynik@gmail.com
Sergiy I. Kovalenko: Zaporizhia State Medical University,
Mayakovsky Ave., 26, Zaporozhia 69035, Ukraine
Svetlana V. Shishkina: SSI Institute for Single Crystals, National
Academy of Sciences of Ukraine, Lenin Ave. 60, Kharkiv 61001,
Ukraine
The structure of compound 3h was additionally deter-
mined by X-ray diffraction study (Figure 1). The tricyclic
fragment is planar within 0.01 Å. The H4…N1 (2.63 Å)
and H16c…N2 (2.54 Å) attractive interactions can be seen
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138ꢀ
ꢀO.Y. Voskoboynik et al.: [1,2,4]Triazino[2,3-c][1,2,3]triazines as antitumor agents
R2
R2
R2
N
N
N
N
NH2
N
N
N2H4 H2O
NaNO2
H
N
O
R3
N
N
R3
N
iPrOH, ∆, 1 h R3
N
AcOH, rt, 2 h
N
O
R1
R1
R1
1a–j
2a–j
3a–j
O
a: R1 = Me; R2 = R3 = H
b: R1 = Ph; R2 = R3 = H
f: R1 = 4-MeO-C6H4; R2 = R3 = H
g: R1 = 4-F-C6H4; R2 = R3 = H
h: R1 = Ph; R2 = Me; R3 = H
i: R1 = Ph; R2 = H; R3 = Br
c: R1 = 4-Me-C6H4; R2 = R3 = H
d: R1 = 4-iPr-C6H4; R2 = R3 = H
e: R1 = 4-tBu-C6H4; R2 = R3 = H
j: R1 = 4-F-C6H4; R2 = H; R3 = Br
Scheme 1ꢀSynthetic route for 3-R1-8-R2-10-R3-2H-benzo[e][1,2,4]triazino[2,3-c][1,2,3]triazin-2-ones 3a–j.
C16
Anticancer assays were performed according to the US NCI
protocol, which was described elsewhere [23–25].
The experimental data show that some of the studied
compounds reveal high antitumor activity. Compound 3d
is the most active with a broad cytostatic activity spectrum.
The range of cancer cells growth for 3d is 13–101% with a
mean growth of 53.7%. Compounds 3b (mean growth 92%,
range of growth, 56–115%), 3c (mean growth 91%, range
of growth, 29–113%) and 3g (mean growth 86%, range
of growth, 18–108%) show moderate anticancer activity.
Compounds 3f and 3h do not show significant antitumor
activity. It should be noted that antitumor activity of syn-
thesized benzo[e][1,2,4]triazino[2,3-c][1,2,3]triazin-2-ones
is selective. Breast cancer cell line MDA-MB-468 is the
most sensitive. Compounds 3b, 3c and 3g inhibit growth
of MDA-MB-468 cells line by 43%, 70% and 81%, respec-
tively. The most active compound 3d shows cytotoxic
activity and reduced initial quantity of MDA-MB-468 cells
at 13%. Compound 3d was evaluated for dose-depended
antitumor activity according to phase 2 of NCI protocol.
Dose-dependency was studied at five concentrations in
10-fold dilution (100–0.01 μm) at 60 lines of nine cancer
cell types at 10-fold dilution (100–0.01 μm, Table 1). As
can be seen, the most sensitive line is breast cancer line
MDA-MB-468. The GI50 value for this line is 0.41 μm.
N2
C7
C8
C6
C5
N3
N4
C3
N5
C4
C2
N1
C11
C10
C12
C13
C9
C1
C15
01
C14
Figure 1ꢀStructure of compound 3h according to X-ray structural
study.
because the van der Waals radii sum is 2.67 Å) [21]. The
phenyl substituent is twisted relatively to the triazine ring
with the C1-C9-C10-C15 torsion angle of -32.6(2)°. This is
probably due by the repulsion between the phenyl group
and the adjacent atoms; the shortened intramolecular
contacts are 2.49 Å for H11…N5 and 2.87 Å for H15…C1
2.86 Å. It can also be suggested that the repulsion between
the phenyl substituent and the triazine moiety promotes
the elongation of the C1-C9 bond to 1.497(1) Å as compared Conclusion
with its mean value of 1.46 Å [22].
The treatment of 3-(2-amino-4-R2-phenyl)-6-R1-1,2,4-tria-
zin-5(2H)-ones with sodium nitrite in acetic acid yielded
3-R1-8-R2-10-R3-2H-benzo[e][1,2,4]triazino[2,3-c][1,2,3]
triazin-2-ones derivatives. Some of the synthesized com-
Anticancer activity
Compounds 3b–d and 3f–h were selected by the pounds exhibit anticancer activity against human tumor
National Cancer Institute (NCI) Developmental Thera- cells, especially breast cancer cell lines. The most active
peutic Program (www.dtp.nci.nih.gov) for the in vitro compound 3d is highly active against breast cancer MDA-
cell line screening to investigate their anticancer activity. MB-468 cell line (GI50ꢀ=ꢀ0.41 μm).
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ꢁꢀꢀꢀ
O.Y. Voskoboynik et al.: [1,2,4]Triazino[2,3-c][1,2,3]triazines as antitumor agentsꢂ
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Table 1ꢀAnticancer activity of compound 3d against individual tumor lines (GI50ꢃ<ꢃ4.00 μm).
Cell line/cancer type
ꢁ
ꢁ
3dꢁ
LC50, μma
Erlotinibb
LC50, μma
ꢁ
GI50, μmaꢁ
TGI, μmaꢁ
GI50, μmaꢁ
TGI, μmaꢁ
Leukemia K-562
ꢃ
ꢃ
3.31ꢃ
1.32ꢃ
3.31ꢃ
3.89ꢃ
3.55ꢃ
3.55ꢃ
2.95ꢃ
2.51ꢃ
2.63ꢃ
3.02ꢃ
0.41ꢃ
ꢃ>ꢃ 100ꢃ
ꢃ>ꢃ 100ꢃ
ꢃ>ꢃ 100ꢃ
ꢃ>ꢃ 100ꢃ
ꢃ>ꢃ 100ꢃ
ꢃ>ꢃ 100ꢃ
ꢃ>ꢃ 100ꢃ
ꢃ>ꢃ 100ꢃ
ꢃ>ꢃ 100ꢃ
ꢃ>ꢃ 100ꢃ
ꢃ>ꢃ 100ꢃ
ꢃ>ꢃ 100ꢃ
ꢃ>ꢃ 100ꢃ
ꢃ>ꢃ 100ꢃ
ꢃ>ꢃ 100ꢃ
ꢃ>ꢃ 100ꢃ
ꢃ>ꢃ 100ꢃ
ꢃ>ꢃ 100ꢃ
ꢃ>ꢃ 100ꢃ
ꢃ>ꢃ 100ꢃ
ꢃ>ꢃ 100ꢃ
ꢃ>ꢃ 100ꢃ
34.4ꢃ
26.24ꢃ
5.83ꢃ
ꢃ>ꢃ 100ꢃ
74.64ꢃ
90.36ꢃ
ꢃ>ꢃ 100ꢃ
ꢃ>ꢃ 100ꢃ
ꢃ>ꢃ 100ꢃ
51.52ꢃ
ꢃ>ꢃ 100ꢃ
ꢃ>ꢃ 100ꢃ
ꢃ>ꢃ 100ꢃ
3.64ꢃ
ꢃ>ꢃ 100
74.64
90.36
ꢃ>ꢃ 100
ꢃ>ꢃ 100
ꢃ>ꢃ 100
51.52
ꢃ>ꢃ 100
ꢃ>ꢃ 100
ꢃ>ꢃ 100
88.71
Leukemia RPMI-8226
Non-small cell lung cancer NCI-H460 ꢃ
Melanoma LOX IMVI
Melanoma UACC-257
Ovarian cancer OVCAR-4
Renal cancer A498
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
5.83ꢃ
97.7ꢃ
10.00ꢃ
2.27ꢃ
Prostate cancer PC-3
Breast cancer MCF7
Breast cancer T-47D
Breast cancer MDA-MB-468
3.64ꢃ
100.0ꢃ
3.97ꢃ
0.14ꢃ
aGI50, Growth inhibition of 50%; TGI, total growth inhibition; LC50, concentration lethal to 50%.
bErlotinib data were obtained from NCI DTP database (https://dtp.cancer.gov).
3-Phenyl-2H-benzo[e][1,2,4]triazino[2,3-c][1,2,3]triazin-2-one
(3b)ꢁThis compound was obtained as white crystalline powder in
93% yield; mp 230–232°C; IR: 1662, 1605, 1584, 1549, 1529, 1479, 1444,
1359, 1333, 1311, 1281, 1244, 1206, 1185, 1160, 1119, 1069, 1058, 1029,
1001, 945, 931, 896, 840, 814, 773, 752, 687, 677, 642, 616 cm-1; 1H NMR:
δ: 8.71 (d, J ꢀ=ꢀ 7.2 Hz, 1H, H-11), 8.45 (d, J ꢀ=ꢀ 7.7 Hz, 1H, H-8), 8.38 (d, J ꢀ=ꢀ
6.5 Hz, 2H, 3-Ph H-2,6), 8.26 (t, J ꢀ=ꢀ 7.7 Hz 1H, H-9), 8.17 (t, 1H, J ꢀ=ꢀ 7.2
Experimental
Melting points were determined in open capillary tubes and were
uncorrected. The elemental analyses (C, H, N, S) were performed
using the ELEMENTAR vario EL Cube analyzer (USA). 1H NMR spectra
(400 MHz) and 13C NMR spectra (100 MHz) were recorded on a Var-
ian-Mercury 400 (Varian Inc., Palo Alto, CA, USA) spectrometer with
TMS as internal standard in DMSO-d6–CCl4 (1:1) mixture. LC-MS were
recorded using a chromatography- mass spectrometric system which
consisted of high performance liquid chromatograph Agilent 1100
Series (Agilent, Palo Alto, CA, USA) equipped with diode-matrix and
mass-selective detector Agilent LC/MSD SL (atmospheric pressure
chemical ionization – APCI). Electron impact mass spectra (EI-MS)
were recorded on a Varian 1200 L instrument at 70 eV (Varian, USA).
Compounds 1a–j and 2a–j were obtained according to the described
synthetic protocols [19, 20].
+
Hz, H-10), 7.72–7.41 (m, 3H, 3-Ph H-3,4,5); EI-MS: m/z 275 (M , 2), 247
(6), 219 (10), 191 (14), 190 (39), 144 (10), 119 (10), 117 (7), 116 (53), 107
(7), 103 (30), 102 (27), 90 (11), 89 (100), 88 (50), 77 (18), 76 (78), 75 (28),
74 (9), 64 (9), 63 (49), 62 (18), 52 (8%); LC-MS: m/z 276. Anal. Calcd for
C15H9N5O: C, 65.45; H, 3.30; N, 25.44. Found: C, 65.47; H, 3.32; N, 25.45.
3-(p-Tolyl)-2H-benzo[e][1,2,4]triazino[2,3-c][1,2,3]triazin-2-one
(3c)ꢁThis compound was obtained as white crystalline powder in
85% yield; mp 235–237°C; 1H NMR: δ 8.69 (d, J ꢀ=ꢀ 7.6 Hz, 1H, H-11), 8.44
(d, J ꢀ=ꢀ 7.7 Hz, 1H, H-8), 8.31 (d, J ꢀ=ꢀ 7.7 Hz, 2H, 3-Ph H-2,6), 8.28- 8.21 (t,
J ꢀ=ꢀ 7.2 Hz, 1H, H-9), 8.17 (t, J ꢀ=ꢀ 7.2 Hz, 1H, H-10), 7.36 (d, J ꢀ=ꢀ 7.6 Hz, 2H, 3
Ph H-3,5), 2.48 (s, 3H, CH3); LC-MS: m/z 290. Anal. Calcd for C16H11N5O:
C, 66.47; H, 3.83; N, 24.21. Found: C, 66.50; H, 3.84; N, 24.26.
General method for the preparation of 3-R1-8-R2-10-R3-
2H-benzo[e][1,2,4]triazino[2,3-c][1,2,3]triazin-2-ones
3a–j
3-(4-Isopropylphenyl)-2H-benzo[e][1,2,4]triazino[2,3-c][1,2,3]
triazin-2-one (3d)ꢁThis compound was obtained as white crystal-
1
line powder in 93% yield; mp 207–209°C; H NMR: δ: 8.70 (d, J ꢀ=ꢀ 7.2
Compound 2a–j (5 mmol) was suspended in acetic acid (20 mL) and
treated with sodium nitrite (0.52 g, 7.5 mmol). The mixture was stirred
at room temperature for 2 h, then cooled, and the resultant precipi-
tate was filtered off, washed by water, dried and crystallized from
acetic acid.
Hz, 1H, H-11), 8.44 (d, J ꢀ=ꢀ 7.3 Hz, 1H, H-8), 8.32 (d, J ꢀ=ꢀ 6.3 Hz, 2H, 3-Ph
H-2,6), 8.25 (t, J ꢀ=ꢀ 6.6 Hz, 1H, H-9), 8.17 (t, J ꢀ=ꢀ 6.2 Hz, 1H, H-10), 7.40 (d,
J ꢀ=ꢀ 6.3 Hz, 2H, 3-Ph H-3,5), 3.05–2.96 (m, 1H, CH(CH3)2), 1.33 (d, J ꢀ=ꢀ 4.3
Hz, 6H, CH(CH3)2); LC-MS: m/z 318. Anal. Calcd for C18H15N5O: C, 68.13;
H, 4.76; N, 22.07. Found: C, 68.17; H, 4.79; N, 22.11.
3-Methyl-2H-benzo[e][1,2,4]triazino[2,3-c][1,2,3]triazin-2-one
(3a)ꢁThis compound was obtained as white crystalline powder in
38% yield; mp 226–228°C; 1H NMR: δ 8.62 (d, J ꢀ=ꢀ 7.8 Hz, 1H, H-11), 8.47
(d, J ꢀ=ꢀ 7.9 Hz, 1H, H-8), 8.26 (t, J ꢀ=ꢀ 7.5 Hz, 1H, H-9), 8.17 (t, J ꢀ=ꢀ 7.5 Hz,
1H, H-10), 2.43 (s, 3H, CH3); 13C NMR: δ 171.9 (C-2), 166.1 (C-11b), 156.0
(C-3), 150.3 (C-7a), 147.3 (C-10), 146.3 (C-9), 140.2 (C-8), 135.3 (C-11),
129.0 (C-11a), 28.9 (CH3), LC-MS: m/z 214. Anal. Calcd for C10H7N5O: C,
56.34; H, 3.31; N, 32.85. Found: C, 56.38; H, 3.35; N, 32.89.
3-(4-(tert-Butyl)phenyl)-2H-benzo[e][1,2,4]triazino[2,3-c][1,2,3]
triazin-2-one (3e)ꢁThis compound was obtained as white crystal-
1
line powder in 83% yield; mp 187–189°C; H NMR: δ: 8.68 (d, J ꢀ=ꢀ 7.7
Hz, 1H, H-11), 8.44 (d, J ꢀ=ꢀ 7.8 Hz, 1H, H-8), 8.31 (d, J ꢀ=ꢀ 7.6 Hz, 2H, 3-Ph
H-2,6), 8.26 (t, J ꢀ=ꢀ 7.4 Hz, 1H, H-9), 8.17 (t, J ꢀ=ꢀ 7.4 Hz, 1H, H-10), 7.56
(d, J ꢀ=ꢀ 7.7 Hz, 2H, 3-Ph H-3,5), 1.40 (s, 9H, C(CH3)3); LC-MS: m/z 332.
Anal. Calcd for C19H17N5O: C, 68.87; H, 5.17; N, 21.13. Found: C, 68.89;
H, 5.21; N, 21.15.
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140ꢀ ꢀO.Y. Voskoboynik et al.: [1,2,4]Triazino[2,3-c][1,2,3]triazines as antitumor agents
3-(4-Methoxyphenyl)-2H-benzo[e][1,2,4]triazino[2,3-c][1,2,3]
triazin-2-one (3f)ꢁThis compound was obtained as white crystal-
line powder in 94% yield; mp 246–248°C; 1H NMR: δ: 8.65 (d, J ꢀ=ꢀ 8.2
Hz, 1H, H-11), 8.50 (d, J ꢀ=ꢀ 7.8 Hz, 1H, H-8), 8.40 (d, J ꢀ=ꢀ 8.6 Hz, 2H, 3-Ph
H-2,6), 8.27 (t, J ꢀ=ꢀ 7.5 Hz, 1H, H-9), 8.19 (t, J ꢀ=ꢀ 7.5 Hz, 1H, H-10), 7.16 (d,
J ꢀ=ꢀ 8.7 Hz, 2H, 3-Ph H-3,5), 3.89 (s, 3H, OCH3); LC-MS: m/z 306. Anal.
Calcd for C16H11N5O2: C, 62.95; H, 3.63; N, 22.94. Found: C, 62.97; H,
3.66; N, 22.95.
X-ray crystallographic study
The colorless crystals 3h (C16H11N5O) are monoclinic. At 193
K, aꢀ=ꢀ7.9168(4), bꢀ=ꢀ11.0274(4), cꢀ=ꢀ15.7059(6) Å, βꢀ=ꢀ102.618(4)°,
Vꢀ=ꢀ1338.0(1) Å3, Mrꢀ=ꢀ289.30, Zꢀ=ꢀ4, space group P21/n, dcalcꢀ=ꢀ1.436 g/cm3,
μ(MoK )ꢀ=ꢀ0.096 mm-1, F(000)ꢀ=ꢀ600. Intensities of 12 976 reflections
(3889 iαndependent, Rintꢀ=ꢀ0.022) were measured on the Xcalibur-3 dif-
fractometer (graphite monochromated MoKα radiation, CCD detector,
ω-scaning, 2Θmaxꢀ=ꢀ60°).
The structure was solved by direct method using SHELXTL
package [26]. Positions of the hydrogen atoms were located from
electron density difference maps and refined by a riding model
with Uisoꢀ=ꢀnUeq (nꢀ=ꢀ1.5 for methyl group and nꢀ=ꢀ1.2 for other hydro-
gen atoms) of the carrier atom. Full-matrix least-squares refinement
against F2 in anisotropic approximation for non-hydrogen atoms
using 3805 reflections was converged to wR2ꢀ=ꢀ0.125 (R1ꢀ=ꢀ0.043 for
2714 reflections with Fꢀ>ꢀ4σ(F), Sꢀ=ꢀ1.025). The final atomic coordi-
nates, and crystallographic data for molecule 3h were deposited to
with the Cambridge Crystallographic Data Centre, 12 Union Road,
CB2 1EZ, UK (fax: +44-1223-336033; e-mail: deposit@ccdc.cam.
ac.uk). They are available on request quoting the deposition num-
ber CCDC1408960).
3-(4-Fluorophenyl)-2H-benzo[e][1,2,4]triazino[2,3-c][1,2,3]tria-
zin-2-one (3g)ꢁThis compound was obtained as white crystalline
1
powder in 88% yield; mp 337–339°C; H NMR: δ: 8.70 (d, J ꢀ=ꢀ 7.8 Hz,
1H, H-11), 8.55–8.37 (m, 3H, H-8, 3-Ph H-2,6), 8.26 (t, J ꢀ=ꢀ 7.1 Hz, 1H,
H-9), 8.18 (t, J ꢀ=ꢀ 7.1 Hz, 1H, H-10), 7.32 (t, J ꢀ=ꢀ 8.5 Hz, 2H, 3-Ph H-3,5);
LC-MS: m/z 294. Anal. Calcd for C15H8FN5O: C, 61.43; H, 2.75; N, 23.88.
Found: C, 61.47; H, 2.76; N, 23.91.
8-Methyl-3-phenyl-2H-benzo[e][1,2,4]triazino[2,3-c][1,2,3]tria-
zin-2-one (3h)ꢁThis compound was obtained as white crystalline
powder in 54% yield; mp 216–218°C; IR: 1663, 1596, 1541, 1484, 1465,
1444, 1364, 1338, 1316, 1273, 1209, 1162, 1087, 1074, 1038, 1011, 906,
845, 814, 805, 771, 756, 695, 676 cm-1; 1H NMR: δ 8.51 (d, J ꢀ=ꢀ 6.9 Hz, 1H,
H-11), 8.37 (d, J ꢀ=ꢀ 7.2 Hz, 2H, 3-Ph H-2,6), 8.12 – 7.95 (m, 2H, H-9,10),
7.58 (m, 3H, 3-Ph H-3,4,5), 2.96 (s, 3H, CH3); 13C NMR: δ 171.1 (C-2), 160.8
(C-11b), 155.4 (C-3), 152.2 (C-7a), 148.2 (C-10), 146.1 (C-9), 142.7 (C-8),
142.7 (3-Ph, C-4), 140.5 (3-Ph, H-3,5), 139.6 (3-Ph, H-2,6), 133.1 (C-11),
128.9 (C-11a), 27.6 (CH3); LC-MS: m/z 290. Anal. Calcd for C16H11N5O: C,
66.43; H, 3.83; N, 24.21. Found: C, 66.44; H, 3.85; N, 24.22.
Acknowledgments: The authors gratefully acknowledge
the Enamine Ltd. (Kiev, Ukraine) for financial support of
this work, and team of the Drug Synthesis and Chemistry
Branch, National Cancer Institute, Bethesda, MD, USA, for
in vitro evaluation of anticancer activity.
10-Bromo-3-phenyl-2H-benzo[e][1,2,4]triazino[2,3-c][1,2,3]tri-
azin-2-one (3i)ꢁThis compound was obtained as white crystalline
1
powder in 82% yield; mp 248–250°C; H NMR: δ: 8.77 (s, 1H, H-11),
8.46–8.24 (m, 1H), 7.66–7.48 (m, 4H, H-8, 9, 3-Ph H-2,6); EI-MS: m/z
+
355 (M , 2), 190 (39), 156 (6), 154 (9), 120 (10), 119 (19), 115 (28), 105 (7),
References
[1] Patel, R.; Kumari, P.; Rajani, D.; Chikhalia, K. Synthesis and
studies of novel 2-(4-cyano-3-trifluoromethylphenylamino)-
4-(quinoline-4-yloxy)-6-(piperazinyl/piperidinyl)-s-triazines
as potential antimicrobial, antimycobacterial and anticancer
agents. Eur. J. Med. Chem. 2011, 46, 4354–4365.
[2] Dahse, F.; Rüttinger, H.; Frohberg, P. Synthesis and
characterization of novel 1,2,4-triazine derivatives
with antiproliferative activity. Bioorg. Med. Chem. 2010, 18,
1816–1821.
[3] Labouta, I.; Eshba, N.; Salama, H. Synthesis of some substituted
triazolo[4,3-b][1,2,4]triazines as potential anticancer agents.
Monatsh. Chem. 1988, 119, 591–596.
[4] Sztanke, K.; Pasternak, K.; Rzymowska, J.; Sztanke, M.;
Kandefer-Szerszen, M.; Dybała, I.; Koziol, A. Identification
of antitumor activity of novel derivatives of 8-aryl-2,6,7,8-
tetrahydroimidazo[2,1-c][1,2,4]triazine-3,4-dione and
8-aryl-4-imino-2,3,7,8-tetrahydroimidazo[2,1-c][1,2,4]triazin-
3(6H)-one. Bioorg. Med. Chem. 2007, 15, 2837–2849.
103 (35), 100 (8), 90 (26), 89 (100), 88 (31), 77 (13), 76 (9), 75 (38), 74
(8), 73 (10), 65 (7), 63 (48), 62 (11), 61 (10), 54 (8), 50 (8); LC-MS: m/z
355. Anal. Calcd for C15H8BrN5O: C, 50.87; H, 2.28; N, 19.77. Found: C,
50.89; H, 2.32; N, 19.81.
10-Bromo-3-(4-fluorophenyl)-2H-benzo[e][1,2,4]triazino[2,3-c]
[1,2,3]triazin-2-one (3j)ꢁThis compound was obtained as white crys-
1
talline powder in 95% yield; mp 237–239°C; H NMR: δ: 8.78 (s, 1H,
H-11), 8.55 – 8.46 (m, 2H, 3-Ph H-2,6), 8.44–8.31 (m, 2H, H-8,9), 7.32 (t,
J ꢀ=ꢀ 8.6 Hz, 2H, 3-Ph H-3,5); LC-MS: m/z 371. Anal. Calcd for C15H7BrFN5O:
C, 48.41; H, 1.90; N, 18.82. Found: C, 48.44; H, 1.93; N, 18.84.
Cytotoxic activity against malignant human tumor cells
Primary anticancer assay was performed at 60 human tumor cell lines
panel derived from nine neoplastic diseases, in accordance with the
protocol of the Drug Evaluation Branch, National Cancer Institute,
Bethesda [23–25]. Three dose response parameters were calculated for
each compound. Growth inhibition of 50% (GI50) – drug concentration [5] Gucký, T.; Řezníčková, E.; Džubák, P.; Hajdúch, M.; Kryštof V.
resulting in a 50% lower net protein increase in the treated as compared
to the net protein increase seen in the control cells, the drug concentra-
tion resulting in total growth inhibition (TGI), and LC50 concentration
Synthesis and anticancer activity of some 1,5-diaryl-3-(3,4,5-
trihydroxyphenyl)-1H-pyrazolo[4,3-e][1,2,4]triazines. Monat.
Chem. 2010, 141, 709–714.
of drug resulting in a 50% reduction in the measured protein at the end [6] Yakhontov, L.; Vakhatova, G. Search for medicinal preparations
of the drug treatment as compared to that at the beginning. The lowest
values were obtained with the most sensitive cell lines.
in the series of 1,3,5-triazines (review). Pharm. Chem. J. 1982,
15, 546–561.
Unauthenticated
Download Date | 6/3/16 10:13 PM

ꢁꢀꢀꢀ
O.Y. Voskoboynik et al.: [1,2,4]Triazino[2,3-c][1,2,3]triazines as antitumor agentsꢂ
ꢂ141
[7] Anderson, E.; Boger, D. Inverse Electron Demand Diels–Alder
Reactions of 1,2,3-Triazines: Pronounced Substituent Effects
on Reactivity and Cycloaddition Scope. J. Am. Chem. Soc. 2011,
133, 12285–12292.
[8] Ohsawa, A.; Arai, H.; Ohnishi, H.; Itoh, T.; Kaihoh, T.; Okada,
M.; Igeta H. Oxidation of 1-aminopyrazoles and synthesis of
1,2,3-triazines. J. Org. Chem. 1985, 50, 5520–5523.
[9] Migawa, M.; Townsend, L. Synthesis and unusual
chemical reactivity of certain novel 4,5-disubstituted
7-benzylpyrrolo[2,3-d][1,2,3]triazines. J. Org. Chem. 2001, 66,
4776–4782.
[10] Thomae, D.; Perspicace, E.; Hesse, S.; Kirsch, G.; Seck, P.
Synthesis of substituted [1,3]thiazolo[4,5-b]pyridines and
[1,3]thiazolo[4,5-d][1,2,3]triazines. Tetrahedron 2008, 64,
9309–9314.
[11] Mohamed, O.; Thabeta, E. Synthesis of some new thieno[2,3-
b]pyridines, pyrido[3′,2′:4,5]-thieno[3,2-d]pyrimidines and
pyrido[3′,2′:4,5]thieno[3,2-d][1,2,3]-triazines. Phosphorus
Sulfur Silicon Relat. Elem. 2000, 166, 149–171.
[16] Kovalenko, S.; Nosulenko, I.; Voskoboynik, A.; Berest, G.;
Antypenko, L.; Antipenko, A.; Katsev A. Novel N-aryl(alkaryl)-2-
[(3-R-2-oxo-2H-[1,2,4]triazino[2,3-c]quinazoline-6-yl)thio]aceta-
mides: synthesis, cytotoxicity, anticancer activity, compare
analysis and docking. Med. Chem. Res. 2013, 22, 2610–2632.
[17] Lima, L.; Barreiro, E. Bioisosterism: a useful strategy for
molecular modification and drug design. Curr. Med. Chem.
2005, 12, 23–49.
[18] Rewcastle, G.; Denny, W.; Bridges, A.; Zhou, H.; Cody, D.;
McMichae, A.; Fry D. Tyrosine Kinase Inhibitors. 5. Synthesis
and Structure-Activity Relationships for 4-[(Phenylmethyl)
amino]- and 4-(Phenylamino)quinazolines as Potent Adeno-
sine 5’-Triphosphate Binding Site. J. Med. Chem. 1996, 38,
3482–3487.
[19] Sergeieva, T.; Voskoboynik, O.; Okovytyy, S.; Kovalenko, S.;
Shishkina, S.; Shishkin, O.; Leszczynski, J. Hydrazinolysis of
3-R-[1,2,4]Triazino[2,3-c]quinazolin-2-ones. Synthetic and
Theoretical Aspects. Phys. Chem. A. 2014, 118, 1895–1905.
[20] Karpenko, O.; Kovalenko, S.; Chekotylo, O.; Shishkina S. A
New One-Step Synthesis of 1,2,4-Triazino[2,3-c]quinazolines.
Heterocycles 2007, 71, 619–626.
[21] Zefirov, Y. Reduced intermolecular contacts and specific inter-
actions in molecular. Crystals. Crystall. Rep. 1997, 42, 865–887.
[22] Burgi, H.-B.; Dunitz, J.D. Structure Correlation; 2nd Edition.
VCH, Weinheim, 1994.
[23] Boyd, M.; Paull, K. Some practical considerations and applica-
tions of the National Cancer Institute in vitro anticancer drug
discovery screen. Drug Dev. Res. 1995, 34, 91–109.
[12] Hunt, J.; Briggs, E.; Clarke, E.; Whittingham, W.; Synthesis and
SAR studies of novel antifungal 1,2,3-triazines. Bioorg. Med.
Chem. Lett. 2007, 17, 5222–5226.
[13] Nosulenko, I.; Voskoboynik, O.; Berest, G.; Safronyuk, S.; Kova-
lenko, S.; Kamyshnyi, O.; Polishchuk, N.; Sinyak, R.; Katsev, A.
Synthesis 3-R-8-R1-9-R2-10-R3-R-6-thioxo-6,7-dihydro-2H-[1,2,4]
triazino[2,3-c]quinazolin-2-ones, its antibacterial and antifun-
gal activity. Sci. Pharm. 2014, 82, 483–500.
[14] Berest, G.; Voskoboynik, A.; Kovalenko, S.; Antypenko, A.;
Nosulenko, I.; Katsev, A.; Shandrovskaya, A. Synthesis and bio- [24] Boyd M. R. The NCI in vitro anticancer drug discovery screen;
logical activity of novel N-cycloalkyl-(cycloalkylaryl)-2-[(3-R-2-
oxo-2H-[1,2,4]triazino[2,3-c]quinazoline-6-yl)thio]acetamides.
Eur. J. Med. Chem. 2011, 46, 6066–6074.
Concept, Implementation and Operation, Cancer Drug Discov-
ery and Development, Vol. 2; Humana Press, 1997.
[25] Monks, A.; Scudiero, D.; Skehan, P.; Shoemaker, R.; Paull, K.;
Vistica, D.; Hose, C.; Langley, J.; Cronise, P.; Vaigro-Wolff, A
Feasibility of a high-flux anticancer drug screen using a diverse
panel of cultured human tumor cell lines. J. Nat. Cancer Inst.
1991, 83, 757–766.
[15] Berest, G.; Voskoboynik, O.; Kovalenko, S.; Nosulenko, I.;
Antypenko, L.; Antypenko, O.; Shvets, V.; Katsev A. Synthesis of
new 6-{[ω-(dialkylamino(heterocyclyl)alkyl]thio}-3-R-2H-[1,2,4]
triazino[2,3-c]quinazoline-2-ones and evaluation of their
anticancer and antimicrobial activities. Sci. Pharm. 2012, 80,
37–65.
[26] Sheldrick, G. A short history of SHELX. Acta Crystallogr. Sect. A
2008, A64, 112–122.
Unauthenticated
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