New Bis-4H-1,2,4-triazoles and Their In Vitro Study as DNA Methylation Inhibitors

Article abstract of DOI:10.1134/S1070363220050047

Abstract: Bis-1,2,4-triazoles with aryl and alkyl linkers were synthesized. In order to increase their hydrophilicity and potential biological activity, pharmacophore electron-deficient groups (carboxylic, carboxamide, nitrile) were introduced into the molecule. Functionalized compounds revealed a new type of biological activity such as inhibition of the level of methylation of tumor DNA.

Full text of DOI:10.1134/S1070363220050047

ISSN 1070-3632, Russian Journal of General Chemistry, 2020, Vol. 90, No. 5, pp. 787–793. © Pleiades Publishing, Ltd., 2020.
Russian Text © The Author(s), 2020, published in Zhurnal Obshchei Khimii, 2020, Vol. 90, No. 5, pp. 681–689.
New Bis-4H-1,2,4-triazoles and Their In Vitro Study
as DNA Methylation Inhibitors
S. V. Dilanyana, T. R. Hovsepyana, L. E. Nersesyana, A. S. Agaronyana, I. S. Danielyana,
N. S. Minasyana, and A. A. Harutyunyana,*
a Scientific and Technological Center of Organic and Pharmaceutical Chemistry of the National Academy of Sciences
of the Republic of Armenia, Yerevan, 0014 Armenia
*e-mail: harutyunyan.artur@yahoo.com
Received December 22, 2019; revised December 22, 2019; accepted December 26, 2019
Abstract—Bis-1,2,4-triazoles with aryl and alkyl linkers were synthesized. In order to increase their hydrophilic-
ity and potential biological activity, pharmacophore electron-deficient groups (carboxylic, carboxamide, nitrile)
were introduced into the molecule. Functionalized compounds revealed a new type of biological activity such as
inhibition of the level of methylation of tumor DNA.
Keywords: bis-1,2,4-triazole, bistiosemicarbazide, linker, cyclization, tumor DNA
DOI: 10.1134/S1070363220050047
Ause of combination of various pharmacophore groups
with a 1,2,4-triazole scaffold in design of macromolecular
drugs resulted in obtaining new structures with diverse
biological activities [1–3]. Recently, we have reported a
new type of biological activity of 1,2,4-triazole derivatives
bearing pharmacologically active alkylsulfanyl fragments
and exhibiting an inhibitory effect on the level of
tumor DNA methylation [4, 5]. DNA methylation
inhibitors are effectively used to treat various types of
cancer [6]. The introduction of electron-withdrawing
groups into the 1,2,4-triazole ring (carboxyl, sulfanyl,
carboxamide, hydroxyl), as well as halogen-substituted
aromatic fragments, contributes to a significant increase
in fungicidal [7, 8], antitumor [9, 10] activity due to a
possible increase stability and bioavailability of these
compounds [11]. The introduction into the structure
of another heterocyclic system [12, 13], the second
1,2,4-triazole ring, as well as various aromatic [14] or
aliphatic [15, 16] linkers leads to bistriazolyl derivatives
and opens up new possibilities in rational drug design.
activity [11], which determined the choice of an aromatic
linker for the synthesis of bis-1,2,4-triazoles 5 and 6.
For the synthesis of bistriazoles with aromatic and
aliphatic linkers 5–8, we used bisthiosemicarbazides
1–4 obtained by the reaction of terephthalic and sebacic
acid bishydrazides with phenyl or allyl isothiocyanates
when boiled in ethanol. The intramolecular cyclization
of thiosemicarbazides 1–4 in an alkaline medium led to
the formation of bis-1,2,4-triazoles 5–8. The obtained
compounds are white crystalline substances that are
poorly soluble in organic solvents and in water. In order
to possibly increase the hydrophilicity and, therefore,
bioavailability of the compounds, as well as to introduce
new pharmacophore groups, compounds 5–8 were
alkylated with some halides containing electron-deficient
groups (2-chloroacetamide, 2-chloroacetic acid, ethyl
chloroacetate) in an aqueous alcohol solution in the
presence of KOH excess. Bis-1,2,4-triazole 7 was also
alkylated with Cl- or F-substituted benzyl chlorides to
afford compounds 17 and 18. To compare the biological
activity of mono- and disubstituted derivatives, we
obtained asymmetric acetamide 21 by reacting bis-1,2,4-
triazole 5 with an equimolar amount of 2- chloroacetamide
(Scheme 2).
Herein, we reported the synthesis of new bis-1,2,4-
triazolyl derivatives 5–27, the structure of which involves
functionalized 1,2,4-triazole fragments linked by aromatic
or alkyl linkers (Scheme 1).
Under similar conditions, we were not able to obtain a
monoalkyl derivative from bis-1,2,4-triazolyl-1,8-octane
7. Using 2-chloroacetamide and 2-chloroacetic acid,
The introduction of various aryl substituents into the
1,2,4-triazole molecule leads to an increase in biological
787

788
DILANYAN et al.
Scheme 1.
S
S
H
H
Lnk
N
N
R1HN
N
H
N
H
NHR1
O
O
1ꢀ4
KOH
R1
R1
Lnk
N
N
SH
SH
N
N
N
N
5ꢀ8
HlgR
R1
R1
R1
N
R1
Lnk
N
N
Lnk
S
N
S
R2H2CS
SCH2R2
N
N
N
N
N
N
N
N
R2
R2
9ꢀ20
22ꢀ27
, R1 = Ph (5), R2 = All (6); Lnk = (CH2)8, R1= Ph (7), R2=All (8).
R1 = Ph, R2=CONH (9), COOH (10); R1 = All, R2=CONH (11),
5ꢀ8, Lnk =
9ꢀ20, Lnk =
,
2
2
Lnk = (CH2)8, R1 = Ph, R2 = CONH2 (14), COOH (15), COOEt (16), 2-ClC6H4 (17),
COOH (12),COOEt (13);
, R1 = Ph,
4-F-C6H4 (18); R1 = All, R2 = CONH2 (19), COOH (20); 22ꢀ27, Lnk =
R2 = CH2CN (22), morpholin-4-yl (23) , R1 =All, R2= CH2CN (24); Lnk = (CH2)8, R1 = Ph, R 2 = CH2CN (25),
(26); R1 = All, R2 = CH2CN (27).
morpholin-4-yl
bis-alkyl derivatives 14 and 15 were prepared with low
yields. Probably, the mobility of the alkyl chain has a
significant effect on the reaction course, which has also
been noted in the study of triazole derivatives with long
chain alkyl groups [17].
compounds 9–13 and 14–20, the singlets of SCH2 groups
at 3.90–3.98 and 3.74–4.35 ppm, respectively, along
with 13C NMR spectroscopy data indicate the occurrence
of alkylation of compounds 5–8 at the thiol group. In
addition, the multiplet and triplet signals of the protons
of the aliphatic linker (CH2)8 at 1.30‒1.45, 1.66‒1.77,
and 2.63 ppm confirm the structure of compounds 14–20.
Chromatographically pure bis-1,2,4-triazoles 9–20 are
stable white crystalline substances.
Structure of bis-1,2,4-triazoles 5–27 was established
by IR, 1H, and 13C NMR spectroscopy methods. In the IR
spectra, there are characteristic absorption bands of the
amide, carboxyl and ester groups, as well as C=C and C=N
bonds in the expected ranges. In the 1H NMR spectra of
RUSSIAN JOURNAL OF GENERAL CHEMISTRY Vol. 90 No. 5 2020

NEW BIS-4H-1,2,4-TRIAZOLES AND THEIR IN VITRO STUDY
789
Scheme 2.
In order to obtain derivatives with a more active lateral
function, we performed cyanoethylation of bistriazoles
5–8 by heating with freshly distilled acrylonitrile in the
presence of a basic triethylamine catalyst (Scheme 1). The
formation of compounds 22, 24, 25, 27 was confirmed by
the appearance in the 1H NMR spectra of triplet signals of
the CH2CN and NCH2 groups in the ranges of 3.13‒3.51
and 4.50 ppm (22, 24), 2.99‒3.03 and 4.14‒4.37 ppm (25,
27), respectively.
doxorubicin by 15.6%. Compound 10 was selected for
further in vivo studies.
In the series of bis-1,2,4-triazoles with octamethylene
linker (CH2), only compound 15 exhibits moderate
activity, which inhibits the level of DNA methylation
by 42.2%. Most active compounds have phenyl group
at position 4 of the triazole ring and a carboxyl group in
the side alkylsulfanyl chain.
In conclusion, new substituted bistriazoles containing
phenyl and octamethylene linkers were synthesized. It
The reaction of bistriazoles 5 and 7 with morpholine in
the presence of formalin (Scheme 1) proceeded at the N‒H
bond of the tautomeric thioxo form of the 1,2,4-triazole
ring, as evidenced by signals of the NCH2N group in
Table 1. The effect of synthesized compounds on the level of
sarcoma-180 DNA methylation
1
the 5.13 and 5.02 ppm regions in the H NMR spectra
Content of bases in DNA,
Methylation
of compounds 28 and 26, respectively. The obtained
Mannich bases are stable white crystalline substances.
mol %
Compound
inhibition, %
–
5-MC±ζ
0.64±0.02
G+C+5-MC
42.66
DNA source
(S-180)
We studied in vitro the effect of bis-1,2,4-triazoles
derivatives obtained on the level of tumor DNAmethyla-
tion on the sarcoma 180 (C-180) model according to the
method reported in [18]. Methylation disorder occurs
in the early stages of the tumor. Unlike mutations that
are irreversible, DNA methylation can be stabilized.
This opens up new opportunities for the early diagnosis
and treatment of cancer. In recent years, a number of
approaches have appeared that allow DNA to be deme-
thylated in cancer, thereby restoring gene expression [19].
Doxorubicin 0.21±0.01
43.40
42.22
42.52
44.10
43.44
40.56
42.78
42.86
42.50
43.42
42.16
44.18
42.72
43.26
45.00
42.92
43.40
43.18
42.68
42.46
43.48
44.64
44.86
41.34
67.2
–
43.8
–
–
–
82.8
31.3
65.6
–
–
42.2
–
–
–
–
–
56.3
34.4
–
37.5
–
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
0.82±0.01
0.36±0.02
1.63±0.01a
1.57±0.02
0.55±0.02
0.11±0.02
0.44±0.01
0.22±0.02
0.70±0.02
0.63±0.01a
0.37±0.01
1.01±0.02
0.79±0.02
0.61±0.01
0.86±0.02
0.55±0.01
0.28±0.01
0.42±0.01
0.73±0.01
0.40±0.01
1.25±0.02a
1.54±0.02
1.01±0.02
Aclear difference between the DNAsamples from the
tumor tissue after exposure to the test compounds was
found only in relation to the amount of 5-methylcytosine
(5-MC) (Table 1). Compound 12 and 21, which inhibit the
level of methylation of tumor DNA by 65.6 and 56.3%,
respectively, have pronounced activity. Compounds
6 (43.8%), 24 (37.5%), 22 (34.4%), and 11 (31.3%)
possess relatively weak demethylating activity. All of
them, along with compounds 12 and 21, are derivatives
of bis-1,2,4-triazoles with an aromatic linker synthesized
from terephthalic acid bishydrazide. The greatest activity
was shown by 2,2'-{benzene-1,4-diylbis[(4-phenyl-4H-
1,2,4-triazole-5,3-diyl)sulfanediyl]}diacetic acid 10,
which inhibits the level of tumor methylation DNA by
82.8%, which exceeds the activity of the control drug
22
23
24
25
26
27
–
–
a p < 0.05.
RUSSIAN JOURNAL OF GENERAL CHEMISTRY Vol. 90 No. 5 2020

790
DILANYAN et al.
5,5'-(Octane-1,8-diyl)bis(4-phenyl-4H-1,2,4-
triazole-3-thiol) (7). Yield 38%, mp 204‒205°С. IR
spectrum, ν, cm–1: 1649 s (C=C), 1570 s (N‒C=S).
1H NMR spectrum, δ, ppm: 0.98‒1.16 m (8H, CH2),
1.31‒1.45 m (4H, CH2), 2.38 t (4H, CH2, J = 7.4 Hz),
7.36‒7.42 m (4HAr), 7.50‒7.59 m (6HAr), 13.63 br. s (2H,
SH). Found, % C 62.17; H 6.26; N 18.31. C24H28N6S2.
Calculated, %: C 62.04; H 6.07; N 18.09.
was found that bistriazoles with a rigid phenyl linker
significantly inhibit the level of DNA methylation in
in vitro experiments on the sarcoma-180 model, while
bistriazoles with a flexible octamethylene linker are less
active.
EXPERIMENTAL
All solvents were purified by distillation before use.
The crystalline starting compounds were recrystallized
from suitable solvents.
5,5'-(Octane-1,8-diyl)bis(4-allyl-4H-1,2,4-triazole-
3-thiol) (8). Yield 90%, mp 173‒175°С. IR spectrum, ν,
cm–1: 3060 w (Н‒C=), 1580 s (C=C), 1500 s (N‒C=S).
1H NMR spectrum, δ, ppm: 1.29‒1.44 m (8H, CH2),
1.64‒1.7 m (4H, CH2), 2.57 t (4H, CH2, J = 7.5 Hz),
4.60 d. t (4H, CH2, J = 5.3, 1.6 Hz), 5.10 d. t. d (2H, =CH,
J = 17.1, 1.6, 1.2 Hz), 5.21 d. t. d (2H, =CH, J = 10.3, 1.6,
1.2 Hz), 5.87 d. d. t (2H, =CH, J = 17.1, 10.3, 5.3 Hz),
13.34 br. s (2H, SH). 13 C NMR spectrum, δС, ppm: 24.5
(2CH2), 25.0 (2CH2), 28.2 (2CH2), 28.3 (2CH2), 44.5
(2NCH2), 116.9 (=CH2), 131.3 (=CH), 151.2 (2C), 166.5
(CS). Found, %: C 55.26; H 7.32; N 21.22. C18H28N6S2.
Calculated, %: C 55.07; H 7.19; N 21.41.
IR spectra were recorded on a NicoletAvatar 330 FT-IR
spectrometer from vaseline oil. 1H and 13C NMR spectra
were recorded on a Varian Mercury-300 instrument from
DMSO-d6 (300 or 75 MHz, respectively), and the internal
standard was TMS. Melting points were determined on a
Boёtius micro-heating table. The reaction progress and
individuality of the obtained compounds were monitored
by TLC on Silicagel 60 F254 plates in dioxane–benzene
(2 : 1) or acetone–benzene (1 : 4) systems, developing
with UV light. Elemental analysis was performed on an
EA 3000 Eurovector automated analyzer.
The starting bisthiosemicarbazides 1–4 were obtained
by boiling a mixture of 0.1 mol of sebacic acid hydrazide
and 0.2 mol of allyl isothiocyanate in 120 mL of ethanol
according to the procedure [20].
S-Alkylation of bis-4H-1,2,4-triazole-3-thiols 5–8
was performed by procedure reported in [21] using 2-fold
molar amount of alkylating agent, the reaction mixture
was heated for 8‒10 h.
General procedure for the synthesis of bis-4H-
1,2,4-triazole-3-thiols 5–8. A mixture of 10 mmol of
thiosemicarbazide 1–4 and 1.68 g (3 mmol) of KOH in
30 mL of water was boiled for 7–8 h. After cooling, the
reaction mixture was filtered, the filtrate was acidified
with glacial acetic acid. The precipitate was filtered
off, washed with water, ethanol and recrystallized from
ethanol.
2,2'-{Benzene-1,4-diylbis[(4-phenyl-4H-1,2,4-
triazole-3,5-diyl)sulfanediyl]}diacetamide (9). Yield
74%, mp >260°C. IR spectrum, ν, cm–1: 3318 br (NH2),
1
1689 s (C=Oamide). H NMR spectrum, δ, ppm: 3.93 s
(4H, SCH2), 7.19 br. s (2H) and 7.63 br. s (2H, CONH2),
7.29 s (4HAr), 7.35‒7.40 m (4HAr), 7.50‒7.57 m (6HAr).
Found, %: C 57.68; H 4.20; N 20.44. C26H22N8O2S2.
Calculated, %: C 57.55; H 4.09; N 20.65.
5,5'-(Benzene-1,4-diyl)bis(4-phenyl-4H-1,2,4-
triazole-3-thiol) (5). Yield 91%, mp >260°С. 1H NMR
spectrum, δ, ppm: 7.31 s (4HAr), 7.37‒7.41 m (4HAr),
7.55‒7.60 m (6HAr), 14.07 br. s (2H, SH). Found, %: C
61.52; H 3.58; N 19.47. C22H16N6S2. Calculated, %: C
61.66; H 3.76; N 19.61.
2,2'-{Benzene-1,4-diylbis[(4-phenyl-4H-1,2,4-
triazole-3,5-diyl)sulfanediyl]}diacetic acid (10). Yield
56%, mp >260°C. IR spectrum, cm–1: 3450 br (O‒H),
1
1731 s (C=O), 1377 s [δ(O‒H)]. H NMR spectrum,
δ, ppm: 3.94 s (4H, CH2), 7.28 s (4HAr), 7.49‒7.56 m
(6HAr), 12.56 br. s (1Н, COOH). Found, %: C 57.53; H
3.56; N 15.51. C26H20N6O4S2. Calculated, %: C57.34;
H 3.70; N 15.43.
5,5'-(Benzene-1,4-diyl)bis(4-allyl-4H-1,2,4,-
triazole-3-thiol) (6). Yield 86%, mp >260°С. IR
spectrum, ν, cm–1: 3040 br (Н‒C=), 1709 s (C=C), 1520 s
(N‒C=S). 1H NMR spectrum, δ, ppm: 4.70‒4.80 m (4H,
NCH2), 4,87 br. d (2H, =CH2, J = 17.0 Hz), 5.13 br. d (2H,
=CH2,J = 10.4 Hz), 5.77‒5.91 m (2H, =CH2), 7.77‒7.89 m
(4HAr), 14.02 br. s (2H, SH). Found, %: C 53.69; H 4.64;
N 23.40. C16H16N6S2. Calculated, %: C 53.91; H 4.52,
N 23.58.
2,2'-{Benzene-1,4-diylbis[(4-allyl-4H-1,2,4-
triazole-3,5-diyl)sulfanediyl]}diacetamide (11). Yield
53%, mp >260°C. IR spectrum, ν, cm‒1: 3363 br (NH2). 1H
NMR spectrum, δ, ppm: 3.95 s (4H, SCH2), 4.68‒4.74 m
(4Н, NCH2), 4.87 d. t. d (2H, =CH2, J = 17.4 Hz),
5.25br. d(2H, =CH2, J=11.6Hz), 5.94‒6.07m(2H, =CH),
7.22 br. s and 7.67 br. s (4H, NH2), 7.77‒7.84 m (4HAr).
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NEW BIS-4H-1,2,4-TRIAZOLES AND THEIR IN VITRO STUDY
791
Found, %: C 51.22; H 4.59; N 23.71. C20H22N8O2S2.
Calculated, %: C 51.05; H 4.71; N 23.81.
(2CH2), 33.9 (2SCH2), 61.1 (2OCH2 ), 127.1 (4CH),
129.9 (6CH), 132.9 (2C), 155.6 (2C), 168.0 (2CO).
Found, %: C 60.24; H 6.55; N 13.10. C32H40N6O4S2.
Calculated, %: C 60.35; H 6.33; N 13.20.
2,2'-{Benzene-1,4-diylbis[(4-allyl-4H-1,2,4-
triazole-3,5-diyl)sulfanediyl]}diacetic acid (12). Yield
53%, mp >260°C. IR spectrum, cm–1: 3420 br (O‒H),
3060 w (Н‒C=), 1717 s (C=O), 1376 s [δ(ОH)]. Found,
%: C 50.72; H 4.40; N 17.62. C20H20N6O4S2. Calculated,
%:C 50.83; H 4.27; N 17.79.
3,3'-(Octane-1,8-diyl)bis[4-phenyl-5-(2-chloro-
benzylsulfanyl)-4H-1,2,4-triazole] (17). Yield 85%, mp
151‒152°C. IR spectrum, ν, cm–1: 3061 w (H‒C=), 1445 s
1
(C=N). H NMR spectrum, δ, ppm: 0.99‒1.14 m (8H,
CH2), 1.35‒1.45 m (4H, CH2), 2.48 t (4H, CH2, J =
7.6 Hz), 4.39 s (4H, SCH2), 7.21‒7.34 m (8HAr),
7.37‒7.44 m (4HAr), 7.49‒7.55 m (6HAr). Found, %: C
64.18; H 5.16; N 11.89. C38H38Cl2N6S2. Calculated, %:
C 64.02; H 5.37; N 11.79.
Diethyl 2,2'-{benzene-1,4-diylbis[(4-allyl-4H-1,2,4-
triazole-3,5-diyl)sulfanediyl]}diacetate (13). Yield
57%, mp >260°C. IR spectrum, ν, cm–1: 3084 w (Н‒C=),
1736 s (C= O). 1H NMR spectrum, δ, ppm: 1.19 t (6H,
CH3, J = 7.1 Hz), 4.12 q (4H, OCH2, J = 7.1 Hz), 4.13 s
(4H, SCH2), 4.68‒4.74 m (4H, NCH2), 4.87 br. d (2H,
=CH2, J = 17.3 Hz), 5.26 br. d (2H, =CH2, J = 10.4 Hz),
5.93‒6.06 m (2H, =CH), 7.28‒7.82 m (4HAr). Found, %:
C 54.32; H 5.24; N 15.99. C24H28N6O4S2. Calculated, %:
C 54.53; H 5.34; N 15.90.
3,3'-(Octane-1,8-diyl)bis[4-phenyl-5-(4-fluoro-
benzylsulfanyl)-4H-1,2,4-triazole (18). Yield 29%,
mp 159‒162°C. IR spectrum, ν, cm–1: 3080 w (Н‒C=),
1600 s (C=C), 1461 s (C=N). 1H NMR spectrum, δ, ppm:
0.98‒1.15 m (8H, CH2), 1.35‒1.46 m (4H, CH2), 2.48 t
(4H, CH2, J = 7.6 Hz), 4.28 s (4H, SCH2), 7.05‒7.13 m
(4HAr), 7.23‒7.36 m (8HAr), 7.50‒7.56 m (6HAr). Found,
%: C 66.72; H 5.92; N 12.20. C38H38F2N6S2. Calculated,
%: C 66.93; H 5.76; N 12.33.
2,2'-{Octane-1,8-diylbis[(4-phenyl-4H-1,2,4-
triazole-3,5-diyl)sulfanediyl]}diacetamide (14). Yield
51%, mp 219‒220°C. IR spectrum, ν, cm–1: 3320 br
1
(NH2), 3091 br (H‒C=), 1693 s (C=Oamide). H NMR
spectrum, δ, ppm: 1.01‒1.17 m (8H, CH2), 1.36‒1.47 m
(4H, CH2), 2.48 t (4H, CH2), 3.84 s (4H, SCH2),
7.18 br. s and 7.62 br. s (4H, NH2), 7.40‒7.46 m (4HAr),
7.55‒7.64 m (6HAr).Found, %: C 61.28; H 6.36; N 20.36.
C28H34N8S2. Calculated, %: C 61.51; H 6.27; N 20.50.
2,2'-{Octane-1,8-diylbis[(4-allyl-4H-1,2,4-triazole-
3,5-diyl)sulfanediyl]}diacetamide (19). Yield 98%, mp
167‒169°C. IR spectrum, ν, cm–1: 3346 br (NH2), 3051 m
1
(Н‒C=), 1600 s (C=O). H NMR spectrum, δ, ppm:
1.31‒1.43 m (8H, CH2), 1.66‒1.77 m (4H, CH2), 2.63 t
(4H, CH2, J = 7.5 Hz), 3.74 s (4H, SCH2), 4.56 d. t (4H,
CH2, J = 4.9, 1.7 Hz), 4.93 d. t. d (2H, =CH, J = 17.1, 1.7,
1.0 Hz), 5.21 d. t. d (2H, =CH, J = 10.4, 1.7, 10.2 Hz),
5.90 d. d. t (2H, =CH, J = 17.1, 10.4, 4.9 Hz), 6.92 br. s
(2H, CONH), 7.51 br (2H, CONH). 13C NMR spectrum,
δС, ppm: 24.1 (2CH2), 26.0 (2CH2), 28.2 (2CH2), 28.3
(2CH2), 36.5 (2SCH2), 45.0 (2NH2), 116.6 (=CH2), 131.7
(=CH),148.5 (2C),154.8 (2С), 168.3 (2CO). Found, %:
C 52.38; H 6.58; N 22.21. C22H34N8O2S2. Calculated, %:
C 52.15; H 6.76; N 22.12.
2,2'-{Octane-1,8-diylbis[(4-phenyl-4H-1,2,4-
triazole-3,5-diyl)sulfanediyl]}diacetic acid (15). Yield
45%, mp 216‒218°C. IR spectrum, cm–1: 3420 br (O‒H),
1
1718 s (C=O), 1377 m [δ(OH)]. H NMR spectrum, δ,
ppm: 0.99‒1.19 m (8H, CH2), 1.35‒1.47 m (4H, CH2),
2.49 t (4H, CH2,J = 7.2 Hz), 3.96 s (4H, CH2), 7.39‒7.46 m
(4HAr), 7.55‒7.62 m (6HAr), 12.94 br. s (2H, COOH).
13C NMR spectrum, δС, ppm: 24.3 (2CH2), 26.1 (2CH2),
28.0 (2CH2), 28.1 (2CH2), 34.4 (2SH2), 127.9 (4CH),
129.9 (4CH), 132.8 (2CH), 199.1 (2C), 155.5 (2C),
169.2 (2CO). Found, %: C 57.70; H 5.67; N 14.28.
C28H32N6O4S2. Calculated, %: C 57.91; H 5.55; N 14.47.
2,2'-{Octane-1,8-diylbis[(4-allyl-4H-1,2,4-triazole-
3,5-diyl)sulfanediyl]}diacetic acid (20). Yield 40%, mp
122‒123°C. IR spectrum, cm–1: 3420 br (O‒H), 1721 s
Diethyl 2,2'-{octane-1,8-diylbis[(4-phenyl-4H-
1,2,4-triazole-3,5-diyl)sulfanediyl]}diacetate (16).
Yield 38%, mp 100‒102°C. IR spectrum, ν, cm–1: 1734 s
1
(C=O), 1381 s [δ(OH)]. H NMR spectrum, δ, ppm:
1.30‒1.45 m (8H, CH2), 1.66‒1.77 m (4H, CH2), 2.63 t
(4H, CH2, J = 7.5 Hz), 3.85 s (4H, SCH2), 4.57 d. t
(4H, CH2, J = 4.8, 1.6 Hz), 4.93 d. t. d (2H, =CH2, J =
17.1, 1.6, 1.0 Hz), 5.22 d. t. d (2H, =CH2, J = 10.5, 1.6,
1.0 Hz), 5.90 d. d. t (2H, =CH, J = 17.1, 10.5, 4.8 Hz),
11.58 br. s (2Н, COOH). 13C NMR spectrum, δС, ppm:
24.1 (2CH2), 26.1 (2CH2), 28.2 (2CH2), 28.3 (2CH2), 35.2
1
(C=O). H NMR spectrum, δ, ppm: 1.00‒1.18 m (8H,
CH2), 1.17 t (6H, CH3, J = 7.0 Hz), 1.36‒1.47 m (4H,
CH2), 2.50 t (4H, CH2, J = 7.2 Hz), 3.98 s (4H, SCH2),
4.08 q (4H, CH2, J = 7.0 Hz), 7.40‒7.45 m (4HAr),
7.56‒7.64 m (6HAr). 13C NMR spectrum, δС, ppm: 13.9
(2CH3), 24.3 (2CH2), 26.0 (2CH2), 27.9 (2CH2), 28.0
RUSSIAN JOURNAL OF GENERAL CHEMISTRY Vol. 90 No. 5 2020

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DILANYAN et al.
(2CH2), 116.7 (2=CH2), 131.6 (2=CH), 147.9 (2C), 154.9
(2C), 168.7 (2CО). Found, %: C 51.71; H 6.45; N 16.27.
C22H32N6O4S2. Calculated, %: C 51.95; H 6.34; N 16.52.
ppm: 16.0 (2CH2), 44.3 (2NHCH2), 47.1 (2NCH2),117.4
(2CH), 118.0 (2=CH2), 127.6 (2C), 129.0 (4CH), 131.2
(2=CH), 149.5 (2C), 167.3 (2CN). Found, %: C 57.35;
H 4.67; N 24.35. C22H22N8S2. Calculated, %: C 57.12;
H 4.39; N 24.22.
2-({5-[4-(5-Sulfanyl-4-phenyl-4H-1,2,4-triazol-3-
yl)phenyl]-4-phenyl-4H-1,2,4-triazol-3-yl}sulfanyl)-
acetamide (21). Yield 72%, mp >260°C. IR spectrum,
ν, cm–1: 3322 br (NH2), 3072 br (Н‒CH=), 1689 s
(C=Oamide). 1H NMR spectrum, δ, ppm: 3.94 s (2H, CH2),
7.14‒7.69 m (16HAr, NH2), 14.00 br. s (1H, SH). Found,
%: C 59.62; H 3.83; N 20.41. C24H19N7OS2. Calculated,
%: C 59.36; H 3.94; N 20.19.
3,3'-[(Octane-1,8-diyl)bis(5-thioxo-4-phenyl-4,5-
dihydro-1H-1,2,4-triazole-3,1-diyl)]dipropanenitrile
(25). Yield 66%, mp 109‒110°C. IR spectrum, ν, cm–1:
2250 m (C≡N). 1H NMR spectrum, δ, ppm: 1.00‒1.18 m
(8H, CH2), 1.34‒1.45 m (4H, CH2), 2.45 t (4H, CH2,
J = 7.5 Hz), 3.09 t (4H, CH2CN, J = 6.4 Hz), 4.41 t (4H,
NCH2, J = 6.4 Hz), 7.40‒7.45 m (4HAr), 7.51‒7.61 m
(6HAr). 13C NMR spectrum, δС, ppm: 16.0 (2CH2), 24.8
(2CH2), 24.9 (2CH2), 27.7 (2CH2), 27.9 (2CH2), 43.9
(2NCH2), 118.0 (4CH), 129.5 (4CH), 129.6 (2CH), 133.9
(2C), 151.3 (2C), 167.1 (2C=S). Found, %: C 64.36; H
4.38; N 19.81. C30H25N8S2. Calculated, %: C 64.15; H
4.49; N 19.95.
N-Cyanoethylation of bis-4H-1,2,4-triazole-3-thiols
5–8. A mixture of 1 mmol of compound 5–8, 3.24 g
(60 mmol) of freshly distilled acrylonitrile, 8 mLof water,
and 6.0 g (60 mmol) of triethylamine was boiled for 10–
12 h. The solution was evaporated, and the crystalline
residue was recrystallized from ethanol.
N-Aminomethylation of bis-4Н-1,2,4-triazole-3-
thiols 5 and 7 was performed by procedure [5] using
2-fold amount of morpholine and paraform.
5,5'-(Octane-1,8-diyl)bis{2-[(morpholin-4-yl)-
methyl]-4-phenyl-2,4-dihydro-1H -1,2,4-triazole-
3,3'-[(Benzene-1,4-diyl)bis(5-thioxo-4-phenyl-4,5-
dihydro-1H-1,2,4-triazole-3,1-diyl)]dipropanenitrile
(22). Yield 57%, mp >260°C. IR spectrum, ν, cm–1: 2249 s
(C≡N). 1H NMR spectrum, δ, ppm: 3.15 t (4H, CH2CN,
J = 6.5 Hz), 4.51 t (4H, NCH2, J = 6.5 Hz), 7.28 s (4HAr),
7.30‒7.37 m (4HAr), 7.46‒7.54 m (6HAr). 13C NMR
spectrum, δС, ppm: 16.0 (CH2CN), 44.4 (NCH2), 118.2
(CN), 126.9, 128.4 (8CH), 129.4 (4CH), 129.7 (2CH),
134.4, 148.5, 168.1. Found, %: C 62.76; H 4.38; N 20.78.
C28H22N8S2. Calculated, %: C 62.90; H 4.15; N 20.46.
1
3-thione} (26). Yield 67%, mp 209‒210°C. H NMR
spectrum, δ, ppm: 1.01‒1.19 m (8H, CH2), 1.33‒1.46 m
(4H, CH2), 2.44 t (4H, CH2, J = 7.4 Hz), 2.68‒2.75 m
[8H, N(CH2)2], 3.53‒3.59 m [8H, O(CH2)2], 7.39‒7.45 m
(4HAr), 7.52‒7.60 m (6HAr). Found, %: C 61.43; H 6.78;
N 16.83. C34H46N8O2S2. Calculated, %: C 61.60; H 6.99;
N 16.90.
3,3'-[(Octane-1,8-diyl)bis(4-allyl-5-thioxo-4,5-
dihydro-1H-1,2,4-triazole-3,1-diyl)]dipropanenitrile
(27).Yield99%,mp67‒69°C.IRspectrum,ν,cm–1:2251m
5,5'-(Benzene-1,4-diyl)bis{2-[(morpholin-4-yl)
methyl]-4-phenyl-2,4-dihydro-1H-1,2,4-triazole-3-
thione} ( 23). Yield 71%, mp >260°C. IR spectrum,
1
(C≡N). H NMR spectrum, δ, ppm: 1.30‒1.45 m (8H,
CH2), 1.66‒1.77 m (4H, CH2), 2.63 t (4H, CH2, J =
7.5 Hz), 2.99 t (4H, CH2CN, J = 6.6 Hz), 4.37 t (4H,
NCH2, J = 6.6 Hz), 4.65 d. t (4H, CH2, J = 5.3, 1.6 Hz),
5.11 d. t. d (2H, = CH2, J = 17.2, 1.6, 1.0 Hz), 5.23 d. t. d
(2H, =CH2, J = 10.4, 1.6, 1.0 Hz), 5.88 d. d. t (2H, =CH,
J = 17.2, 10,4, 5.3 Hz). 13C NMR spectrum, δC, ppm:
15.7 (2CH2), 24.3 (2CH2), 24.9 (CH2), 28.0 (2CH2),
28.3 (2CH2), 43.8 (2NCH2), 45.7 (2NCH2), 116.4 (2CN),
117.3 (2=CH2), 130.7 (2=CH), 151.1 (2C), 166.2 (2C=S).
Found, %: C 57.66; H 6.95; N 22.61. C24H34N8S2.
Calculated, %: C 57.80; H 6.87; N 22.47.
1
ν, cm–1: 3071 ср (H‒C=). H NMR spectrum, δ, ppm:
2.72‒2.83 m [8H, N(CH2)2], 3.54‒3.63 m [8H, O(CH2)2],
5.14 s (4H, NCH2N), 7.28 s (4HAr), 7.32‒7.38 m (4HAr),
7.43‒7.51 m (6HAr). Found, %: C 61.44; H 5.33; N 17.67.
C32H34N8O2S2. Calculated, %: C 61.32; H 5.46; N 17.88.
3,3'-[(Benzene-1,4-diyl)bis(4-allyl-5-thioxo-4,5-
dihydro-1H-1,2,4-triazole-3,1-diyl]dipropanenitrile
(24). Yield 98%, mp >260°C. IR spectrum, ν, cm–1: 2252 s
1
(C≡N). H NMR spectrum, δ, ppm: 3.11‒3.18 m (4H,
CH2, NCH2), 4.47‒4.54 m (4H, CH2CN), 4.79‒4.85 m
(4H, NCH2), 4.90 br. d (2H, =CH2, J = 17.4 Hz),
5.16 br. d (2H, =CH2, J = 10.6 Hz), 5.79‒5.93 m (2H,
=CH), 7.87‒7.94 m (4HAr). 13C NMR spectrum, δС,
CONFLICT OF INTEREST
No conflict of interest was declared by the authors.
RUSSIAN JOURNAL OF GENERAL CHEMISTRY Vol. 90 No. 5 2020

NEW BIS-4H-1,2,4-TRIAZOLES AND THEIR IN VITRO STUDY
793
Sun, P., Zhany, Y.J., Yang, X.H., Jiang, Y.Y., and
Zhang, D.Z., Bioorg. Med. Chem. Lett., 2008, vol. 18,
p. 3261.
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