Synthesis and Antioxidant Activity of New Neomenthyl and Caranyl Thiotriazoles

Article abstract of DOI:10.1007/s10600-018-2504-y

New neomenthyl and caranyl thiotriazoles were synthesized in up to 80% yields. Studies of the antioxidant and membrane-protective activity of the synthesized compounds in vitro showed that sulfides with a carane moiety were more active than analogs with a

Full text of DOI:10.1007/s10600-018-2504-y

DOI 10.1007/s10600-018-2504-y
Chemistry of Natural Compounds, Vol. 54, No. 5, September, 2018
SYNTHESIS AND ANTIOXIDANT ACTIVITY OF NEW
NEOMENTHYL AND CARANYL THIOTRIAZOLES
1
1*
2
Yu. V. Gyrdymova, D. V. Sudarikov, O. G. Shevchenko,
1
3
1
4
S. A. Rubtsova, P. A. Slepukhin, S. A. Patov, F. A. Lakhvich,
F. S. Pashkovskii, and A. V. Kuchin
4
1
New neomenthyl and caranyl thiotriazoles were synthesized in up to 80% yields. Studies of the antioxidant
and membrane-protective activity of the synthesized compounds in vitro showed that sulfides with a carane
moiety were more active than analogs with a neomenthane substituent.
Keywords: triazoles, neomenthane and carane sulfides, antioxidant and membrane-protective activity, erythrocytes.
The high biological activity of 1,2,4-triazoles and their derivatives has stimulated interest in them over the last few
decades. For example, 1,2,4-triazoles exhibited anti-inflammatory [1, 2], sedative, and antibacterial properties [3–6] and
antifungal [7, 8], antitumor [2], and antiviral [9–11] activity.
Sulfur (S)-containing 1,2,4-triazoles exhibited antioxidant and antiradical activity. Both the thiol and thione forms
were active with the latter more reactive toward DPPH than the former because of the electronically unsaturated C=S bond that
was involved in inhibiting peroxide radicals [12, 13].
Thio derivatives of menthane, bornane, and isobornane monoterpenoids also exhibited broad spectra of biological
activity, with the antioxidant activity especially prominent [14, 15].
Previously, cis- and trans-myrtanyl thiotriazoles with antioxidant and membrane-protective activity were prepared
by us [16]. In continuation of that work, we synthesized the series of monoterpene sulfides 5–10 with neomenthane and
carane moieties and studied their antioxidant properties. Myrtanyl thiotriazoles with 1,5-diphenyl-1,2,4-triazole fragments
were highly cytotoxic [16] so that derivatives with this substituent were replaced by monoterpene 3-amino-1,2,4-triazoles.
10
4
1
2
R
S
OTs
OH
OH
7
8
4
TsCl, Py
RSH, a,b
1
3
5 - 7
10
N
N
N
N
N N
S
OTs
3
R
R =
NH
N
H
N
N
H
2
1
4
8 - 10
5, 8
6, 9
7, 10
8
2
a. Cs CO –TBAI, EtOH (DMF), 60°C; b. NaH, DMF, 60°C
2
3
Scheme 1
1) Institute of Chemistry, Komi Scientific Center, Ural Branch, Russian Academy of Sciences, 48 Pervomaiskaya,
Syktyvkar, 167000; e-mail: sudarikov-dv@chemi.komisc.ru; 2) Institute of Biology, Komi Scientific Center, Ural Branch,
RussianAcademy of Sciences, 28 Kommunisticheskaya, Syktyvkar, 167982; 3) I. Ya. Postovsky Institute of Organic Synthesis,
Ural Branch, RussianAcademy of Sciences, 22 S. Kovalevskoi St., Ekaterinburg, 620137; 4) Institute of Bioorganic Chemistry,
National Academy of Sciences of Belarus, 5/2 Akad. Kuprevicha St., Minsk, Republic of Belarus, 220141. Translated from
Khimiya Prirodnykh Soedinenii, No. 5, September–October, 2018, pp. 750–754. Original article submitted March 16, 2018.
©
0009-3130/18/5405-0883 2018 Springer Science+Business Media, LLC
883

C16
C10
C15
C9
C8
C11
C12
C14
N1
C5
N4
S1
C7
C2
N3
N5
C13
Fig. 1. X-ray structure of sulfide 10.
Monoterpenyl thiotriazoles 5–10 were prepared from L-menthol (1) and 4-isocaranol (2) through the corresponding
tosylates 3 and 4 (Scheme 1). Sulfides 5–10 formed via a bimolecular nucleophilic substitution with full configuration inversion
at the adjoining chiral C atom of the monoterpene. This was demonstrated previously by us using formation of monoterpenyl
imidazoles as examples [17, 18].
1
2
1
8
NMR spectra contained resonances for the terpene and aromatic components. Cross peaks H –H and H –H in
1
1
4
3
4
6
H– H NOESY spectra of 5–7 and H –H and H –H , of 8–10, confirmed the configuration of the neomenthane and carane
moieties, respectively. IR spectra showed characteristic absorption bands for triazole rings at 3110 (N–H), 1600–1400 (C=N),
–1
and 1120 cm (N–N).
An X-ray crystal structure analysis (XSA) found that two crystallographically independent molecules of 10 crystallized
as a hemihydrate in a chiral orthorhombic space group. Figure 1 shows a general view of the molecule with the experimental
atomic numbering. Atoms of the second molecule had the additional index “A”. The molecules differed by the mutual
orientation of the caranyl fragment and the heterocycle. The torsion angles were C(2)S(1)C(7)C(8) = –89.0(3)° and
C(2A)S(1A)C(7A)C(8A) = 176.6(2)°. The other molecular characteristics were similar to each other because of the
conformational rigidity of the structural framework in the sulfide substituents.
The crystal packing was determined by an extended three-dimensional system of intermolecular H-bonds involving
H O, amines, and heterocyclic N atoms.
2
The biological activity of monoterpenyl thiotriazoles 5–10 was assessed in cell (laboratory mouse blood erythrocytes)
and acellular (laboratory mouse brain lipids) models. All tested compounds exhibited high antioxidant activity (AOA) in the
acellular model at a concentration of 1 μM. Sulfides 7 and 10 with amines on the triazole ring turned out to be the most active
(Fig. 2). Previously, 2-aminophenyl(4,5-epoxycaryophyll-9-ylmethyl)disulfide, which also contained an amine in its structure,
displayed high AOA in this model in our hands [19]. The AOA decreased considerably if the monoterpenyl thiotriazole
concentration was reduced to 100 μM (Fig. 2).
The insignificant cytotoxicity (hemolytic activity) of the sulfides that was found at a concentration of 100 μM facilitated
further research on the antioxidant and membrane-protective activity using blood erythrocytes as a model. Table 1 presents
the analytical results.
Compounds 7 and 10, which contained amines, showed statistically significant pro-oxidant activity according to an
analysis of the activities of 5–10 at concentrations of 10 and 100 μM for mammal blood erythrocytes. This was indicated by
the increased death of erythrocytes affected by H O and the increased conversion of oxyhemoglobin into methemoglobin. In
2
2
addition, sulfides 5, 6, 8, and 9 without amines on the triazole ring had moderate membrane-protective and antioxidant activity
at both indicated concentrations, although they were inferior in this respect to the reference drug ionol (BHT). Caranyl
thiotriazoles 8 and 9 were the most active derivatives synthesized by us. They at a concentration of 100 μM not only protected
effectively cells from destruction but also prevented oxidation of native hemoglobin and accumulation of secondary lipid
peroxidation (LPO) products. The pro-oxidant activity of 7 and 10 disappeared if the concentration was lowered. Thus, these
compounds at a concentration of 1 μM exhibited AOA comparable to the protective effect of the other tested compounds
(Table 1).
Therefore, sulfides with a caranyl moiety were more active than those with a neomenthyl substituent under otherwise
equal conditions. Compounds 7 and 10 with amines on the triazole ring had the greatest AOA. However, these compounds at
high concentrations (10 and 100 μM) in the H O -induced hemolysis model showed significant pro-oxidant activity that could
2
2
only be avoided by reducing the concentration to 1 μM. Conversely, the membrane-protective and antioxidant activity of
sulfides without amines (5, 6, 8, and 9) increased with increasing concentration and did not manifest pro-oxidant activity.
884

TABLE 1. Assessment of ComparativeAntioxidant and Membrane-protectiveActivity of Neomenthyl and Caranyl Thiotriazoles
in a Cell Model
Conditions
Control
ÂÍÒ
5
6
7
8
9
10
Concentration 100 M
μ
1
2
3
–
–
–
38.2 1.1
0.96 0.08
2.73 0.06
22.3 0.6
0.94 0.03
2.22 0.08
22.0 0.8
0.50 0.09
2.56 0.07
50.8 2.7
1.11 0.16
2.08 0.06
11.8 0.5
0.67 0.07
1.49 0.13
12.0 0.5
0.63 0.05
1.67 0.15
49.1 3.4
1.13 0.15
2.38 0.08
Concentration 10 M
μ
1
2
3
39.4 1.8
0.77 0.06
2.43 0.03
28.7 1.6
0.55 0.04
2.03 0.05
28.8 0.9
0.48 0.05
2.06 0.04
54.1 1.0
1.11 0.06
2.53 0.08
34.1 0.9
0.72 0.02
2.19 0.08
30.8 1.0
0.67 0.06
2.19 0.08
62.4 2.5
1.43 0.11
2.68 0.04
8.2 0.5
0.64 0.05
1.87 0.04
Concentration 1 M
μ
1
2
3
–
–
–
–
–
–
46.30 1.10
0.79 0.03
2.78 0.03
35.3 0.8
0.65 0.06
2.26 0.09
40.5 1.4
0.82 0.02
2.36 0.03
36.9 0.1
0.76 0.05
2.63 0.15
37.9 0.8
0.66 0.04
2.24 0.08
13.0 0.6
0.55 0.03
1.98 0.07
______
1) Membrane-protective activity (MPA, %), degree of erythrocyte hemolysis by H O (incubation for 5 h); 2) ratio of oxidized
2
2
(methemoglobin) and native (oxyhemoglobin) forms in erythrocytes affected by H O (metHb/oxyHb); 3) concentration of
2
2
secondary LPO products in suspension of erythrocytes affected by H O (TBA–AP, nmol/mL).
2
2
90
80
70
60
50
40
30
20
10
0
C
I
5
6
7
8
9
10
- 100 μM
- 1
μ
M
Fig. 2. Effects of 5–10 at concentrations of 100 μM and 1 μM on
TBA–AP contents in brain homogenate 1 h after inducing LPO
2+
(ascorbate/Fe initiator); C, control; I, intact sample.
EXPERIMENTAL
IR spectra were recorded from thin layers or KBr pellets on a Shimadzu IR Prestige 21 FT-IR spectrometer. PMR and
13
C NMR spectra were recorded in CDCl and MeOD with solvent resonances as internal standards on a Bruker Avance-300
3
1
13
13
spectrometer (300.17 MHz for H and 75.48 MHz for C). C NMR spectra were recorded in J-modulation mode. Resonances
in PMR and C NMR spectra were fully assigned using two-dimensional homo- ( H– H COSY, H– H NOESY) and
13
1
1
1
1
1
13
1
13
heteronuclear experiments ( H– C HSQC, H– C HMBC). Mass spectra were measured in a Thermo Finnigan LCQ Fleet
instrument equipped with an MS detector scanning mass range m/z 50–2000 (ESI, 40 eV). Melting points were determined on
a Sanyo Gallenkamp melting point apparatus (MPD.350.BM3.5) and were uncorrected. Optical rotation angles were measured
on a PolAAr 3001 automatic digital polarimeter (England). The XSA used an Xcalibur 3 automated X-ray diffractometer
according to the standard procedure ([Mo Kα-radiation, graphite monochromator, 295(2) K, ω-scanning in 1° steps]. Absorption
corrections were applied empirically. The structure was solved and refined using the Olex2 software [20]. The structure was
solved using the Charge Flipping method of the Superflip program [21] and was refined by the SHELXL program using
2
anisotropic least-squares methods over F for non-hydrogen atoms [22]. H atoms of C–H groups were added at the calculated
positions and included in the refinement as a rider model. Amine and water protons were refined isotropically and independently.
885

TLC used Sorbfil plates and phosphomolybdic acid solution detector. Column chromatography used Alfa Aesar silica gel
(0.06–0.2 mm). Solvents were used without further purification.
The sulfides were synthesized using L-menthol, 3-mercapto-1,2,4-triazole, 3-mercapto-4-methyl-1,2,4-triazole, and
3-mercapto-5-amino-1,2,4-triazole (Alfa Aesar).
Isocaranol 2 was prepared from 3-carene by the published method [23].
Tosylates of menthol (3) and isocaranol (4) were synthesized by the literature methods [24]. Antioxidant activity of
the sulfides was estimated in vitro from the ability to inhibit LPO in laboratory mouse brain lipids [25, 26]. Excised brain was
homogenized (10%) in normal saline (pH 7.4) and centrifuged for 10 min. The supernatant (S1) containing water, proteins,
DNA, RNA, and lipids (cholesterol, galactolipids, individual phospholipids, and gangliosides) was collected [25, 27].
The tested compounds were added to the supernatant as EtOH solutions (final concentrations 100 μM and 1 μM). LPO was
initiated after 30 min by adding freshly prepared FeCl (3 μM) and ascorbic acid (300 μM) [28]. Then, the test samples were
2
incubated for 1 h at 37°C with slow stirring. The contents of secondary LPO products reacting with 2-thiobarbituric acid
(TBA-AP) were determined using a Thermo Spectronic Genesys 20 spectrophotometer (USA) at 532 nm. An extinction
5
–1
–1
coefficient of 1.56 × 10 M cm was used for the calculations [26, 29].
Toxicity and antioxidant and membrane-protective activity of the compounds were determined using a suspension
(0.5%) of laboratory mouse erythrocytes in phosphate-buffered saline (PBS, pH 7.4). The tested compounds were dissolved
beforehand in EtOH. Toxicity (hemolytic activity) was assessed from the degree of erythrocyte hemolysis 1–5 h after adding
the solutions of the tested compounds.
MPA and AOA were determined from the degree of inhibition of H O -induced hemolysis and slowed accumulation
2
2
of secondary LPO products and oxidation of oxyhemoglobin in erythrocytes. For this, hemolysis was initiated 30 min after
adding solutions of the tested compounds (final concentrations 1, 10, and 100 μM) to the erythrocyte suspension by adding
H O (0.006%). Ionol (BHT) was used as a reference standard. Then, the reaction mixture was incubated in a thermostatted
2
2
shaker with slow stirring at 37°C for 5 h. An aliquot of the incubation medium was taken every hour and centrifuged for 5 min
(1600 g). The degree of hemolysis was determined from the hemoglobin content in the supernatant using a Thermo Spectronic
Genesys 20 spectrophotometer (USA) at 524 nm [30]. The percent hemolysis was calculated relative to total hemolysis of the
sample. The content of secondary LPO products reacting with 2-thiobarbituric acid (TBA–AP) was determined as indicated
above. Accumulation of hemoglobin oxidation products was evaluated by analyzing absorption spectra in the range 540–640 nm.
The contents of native and oxidized hemoglobin (oxyHb and metHb) were calculated considering the corresponding extinction
coefficients [31]. Each experiment was performed in 4–6 replicates. Results were processes statistically and figures were
drawn using Microsoft Office Excel 2007 software.
General Method for Synthesizing Thio Derivatives 5, 6, 8–10. Thiol (1.2 mmol) was dissolved in EtOH (5 mL,
DMF for 10), treated with Cs CO (1.2 mmol) and TBAI (1.2 mmol), stirred for 5 min, treated with a solution of the tosylate
2
3
(1 mmol), and stirred and heated (60°C). The reaction was monitored by TLC. When the reaction was finished, the solvent
was evaporated. The residue was extracted with EtOAc. The extract was washed with saturated NaCl solution. The combined
organic fractions were dried over Na SO . The target sulfides were isolated by column chromatography over silica gel.
2
4
3-{[(1S,2S,5R)-2-Isopropyl-5-methylcyclohexyl]thio}-4H-1,2,4-triazole (5). White powder, yield 29%, mp 140°Ñ,
23
–1
R 0.22 (CHCl ), [α] + 77.5° (c 0.4, CÍCl ). IR spectrum (KBr, ν, cm ): 3136, 3113 (N–H), 1568, 1348 (C=N), 1261
f
3
D
3
– 1
(N–N). Mass spectrum (ESI, 40 eV), m/z (I , %): 238.23 (100) [M – Í] . H NMR spectrum (300 MHz, CDCl , δ, ppm, J/Hz):
rel
3
0.89 (3Í, d, J = 6.5, Ìå-7), 0.93 (6Í, d, J = 6.5, Ìå-9, 10), 0.98–1.06 (1Í, m, Í-4α), 1.10 (1Í, dd, J = 12.6, 2.6, Í-3α),
1.15–1.28 (1Í, m, Í-2), 1.28–1.41 (1Í, m, Í-6α), 1.54–1.71 (1Í, m, Í-5), 1.72–1.98 (3Í, m, Í-3β, 4β, 8), 2.04 (1Í, dd,
13
J = 13.6, 2.2, Í-6β), 4.30 (1Í, s, Í-1), 8.20 (1Í, s, Í-14), 10.10 (1Í, br.s, NÍ). C NMR spectrum (75 MHz, CDCl , δ,
3
ppm): 20.56 (Ñ-9), 20.98 (Ñ-10), 21.96 (Ñ-7), 26.53 (Ñ-3), 27.13 (Ñ-8), 35.09 (Ñ-4), 41.39 (Ñ-6), 48.39 (Ñ-2), 50.04 (Ñ-1),
148.30 (Ñ-14), 156.55 (Ñ-11).
3-{[(1S,2S,5R)-2-Isopropyl-5-methylcyclohexyl]thio}-4-methyl-4H-1,2,4-triazole (6). White powder, yield 51%,
27
–1
mp 82°Ñ, R 0.20 (CHCl –iPrOH, 50:2), [α] + 78.7° (c 0.8, CÍCl ). IR spectrum (KBr, ν, cm ): 3257 (N–H), 1558 (C=N),
f
3
D
3
+ 1
1278 (N–N). Mass spectrum (ESI, 40 eV), m/z (I , %) 254.08 (53) [M + Í] . H NMR spectrum (300 MHz, CDCl , δ, ppm,
rel
3
J/Hz): 0.86 (3Í, d, J = 6.5, Ìå-7), 0.81 (3Í, d, J = 6.5, Ìå-9), 0.85 (3Í, d, J = 6.2, Ìå-10), 0.95–1.02 (1Í, m, Í-4β),
1.03–1.41 (3Í, m, Í-2, 3β, 6β), 1.52–1.68 (1Í, m, Í-5), 1.71–1.96 (3Í, m, H-3α, 4β, 8), 2.09 (1Í, dd, J = 14.1, 2.6, Í-6α), 3.57
13
(3Í, s, Ìå-16), 4.41 (1Í, br.s, Í-1), 8.14 (1Í, s, Í-14). C NMR spectrum (75 MHz, CDCl , δ, ppm): 20.91 (Ñ-9), 21.30
3
(Ñ-10), 21.20 (Ñ-7), 25.16 (Ñ-3), 25.48 (Ñ-8), 29.05 (Ñ-5), 32.65 (Ñ-16), 35.07 (Ñ-4), 39.12 (Ñ-6), 46.54 (Ñ-2), 50.7 (Ñ-1),
144.7 (Ñ-14), 151.3 (Ñ-11).
886

3-{[(1S,3S,4R,6R)-4,7,7-Trimethylbicyclo[4.1.0]heptan-3-yl]thio}-4H-1,2,4-triazole (8). Colorless liquid, yield
23
–1
52%, R 0.22 (CHCl –iPrOH, 50:2), [α] + 18.3° (c 0.1, CHCl ). IR spectrum (KBr, ν, cm ): 3105 (NH), 1514, 1332 (C=N),
f
3
D
3
+ 1
1271 (N–N). Mass spectrum (ESI, 40 eV), m/z (I , %) 238.13 (80) [M + Í] . H NMR spectrum (300 MHz, CDCl , δ, ppm, J/Hz):
rel
3
0.57 (1Í, td, J = 8.8, 5.6, Í-6), 0.71 (1Í, q, J = 8.2, Í-1), 0.84 (1Í, m, Í-2α), 0.95 (3Í, d, J = 6.8, Ìå-10), 0.97 (3Í, s,
Ìå-8), 0.99 (3Í, s, Ìå-9), 1.43 (1Í, m, Í-5α), 1.84 (1Í, ddd, J = 14.4, 8.7, 5.4, Í-2β), 2.0 (1Í, m, Í-3), 2.32 (1Í, dt,
13
J = 15.3, 7.7, Í-5), 4.0 (1Í, q, J = 6.9, Í-4), 8.22 (1Í, s, Í-14), 11.78 (1Í, br.s, NH). C NMR spectrum (75 MHz, CDCl ,
3
δ, ppm): 15.7 (Ñ-9), 17.8 (Ñ-7), 18.8 (Ñ-10), 20.3 (Ñ-6), 21.4 (Ñ-1), 25.2 (Ñ-5), 25.8 (Ñ-2), 28.4 (Ñ-8), 30.7 (Ñ-3), 48.2 (Ñ-4),
147.8 (Ñ-14), 156.7 (Ñ-11).
4-Methyl-3-{[(1S,3S,4R,6R)-4,7,7-trimethylbicyclo[4.1.0]heptan-3-yl]thio}-4H-1,2,4-triazole (9). Yellowish
24
–1
powder, yield 80%, mp 58°Ñ, R 0.11 (EtOAc), [α] +22.1° (c 0.5, CHCl ). IR spectrum (KBr, ν, cm ): 3072, 1512 (C=N),
f
D
3
+ 1
1215 (N–N). Mass spectrum (ESI, 40 eV), m/z (I , %) 252.21 (100) [M + H] . H NMR spectrum (300 MHz, CDCl , δ, ppm, J/Hz):
rel
3
0.62 (1Í, td, J = 8.8, 5.6, Í-6), 0.75 (1Í, q, J = 8.2, Í-1), 0.85 (1Í, m, Í-2α), 0.97 (3Í, d, J = 5.5, Ìå-10), 0.98 (3Í, s,
Ìå-8), 1.01 (3Í, s, Ìå-9), 1.47 (1Í, m, Í-5α), 1.87 (1Í, m, Í-2β), 2.40 (1Í, m, Í-3), 3.59 (3Í, s, Ìå-16), 4.16 (1Í, q,
13
J = 6.8, Í-4), 8.33 (1Í, s, Í-14). C NMR spectrum (75 MHz, CDCl , δ, ppm): 15.8 (Ñ-9), 17.8 (Ñ-7), 18.8 (Ñ-10), 20.2
3
(Ñ-6), 21.4 (Ñ-1), 25.3 (Ñ-5), 25.9 (Ñ-2), 28.4 (Ñ-8), 30.6 (Ñ-3), 31.0 (Ñ-16), 48.2 (Ñ-4), 144.7 (Ñ-14), 151.9 (Ñ-11).
5-{[(1S,3S,4R,6R)-4,7,7-Trimethylbicyclo[4.1.0]heptan-3-yl]thio}-4H-1,2,4-triazolyl-3-amine (10). White powder,
24
–1
yield 57%, mp 139°Ñ, R 0.31 (petroleum ether–i-PrOH, 7:1), [α] +35.6° (c 0.3, EtOH). IR spectrum (KBr, ν, cm ): 3414
f
D
+ 1
(NH ), 3192 (NH), 1649, 1496 (C=N), 1296 (N–N). Mass spectrum (ESI, 40 eV), m/z (I , %) 252.21 (100) [M] . H NMR
2
rel
spectrum (300 MHz, MeOD-d , δ, ppm, J/Hz): 0.56 (1Í, td, J = 8.8, 5.6, Í-6), 0.75 (1Í, q, J = 8.2, Í-1), 0.84 (1Í, m, Í-2α),
4
0.94 (3Í, d, J = 5.5, Ìå-10), 0.95 (3Í, s, Ìå-8), 0.97 (3Í, s, Ìå-9), 1.33–1.48 (1Í, m, Í-5α), 1.71–1.85 (1Í, m, Í-2β),
13
1.85–1.89 (1Í, m, Í-3), 2.18–2.34 (1Í, m, Í-2β), 3.77 (1Í, q, J = 6.8, Í-4), 4.90 (2Í, br.s, NH ). C NMR spectrum
2
(75 MHz, MeOD-d , δ, ppm): 16.4 (Ñ-9), 18.8 (Ñ-7), 19.4 (Ñ-10), 21.5 (Ñ-6), 23.0 (Ñ-1), 26.2 (Ñ-5), 26.9 (Ñ-2), 29.1 (Ñ-8),
4
32.4 (Ñ-3), 47.9 (Ñ-4), 146.7 (Ñ-14), 170.9 (Ñ-11).
°
XSA results: C H N O S, MM = 261.39; orthorhombic system, a = 9.9681(7), b = 9.9931(8), c = 27.9057(19) A;
12 21
4 0.5
° 3
–1
V = 2779.7(4) A ; 295(2) K; space group P2 2 2 ; Z = 8; μ(Mo Kα) = 0.224 mm . A total of 15,467 reflections, of which
1 1 1
7,426 were independent, were collected (R = 0.0654). The final refinement parameters were wR = 0.1971 (all reflections)
int
2
and R = 0.0707 [I > 2σ(I)].
1
5-{[(1S,2S,5R)-2-Isopropyl-5-methylcyclohexyl]thio}-4H-1,2,4-triazolyl-3-amine (7). Menthol tosylate (300 mg,
0.97 mmol) and 3-amino-1,2,4-triazolyl-3-thiol (137 mg, 1.16 mmol) were dissolved in DMF (5 mL), treated with NaH
(49 mg, 1.2 mmol, 57%), and reacted at 60°C. The course of the reaction was monitored by TLC. When the reaction was
finished, the solvent was vacuum distilled (65°C, 10 mm Hg). The residue was extracted with EtOAc. The extract was washed
with NaCl solution and dried over Na SO . The product was isolated by column chromatography over silica gel with elution
2
4
25
by CHCl –i-PrOH (10:1). White powder, yield 50%, mp 177–178°Ñ, R 0.16 (CHCl –iPrOH 10:1), [α] +91.7° (c 0.2,
EtOH). IR spectrum (KBr, ν, cm ): 3460 (NH ), 3160 (N–H), 3124, 1500 (C=N), 1269 (N–N). Mass spectrum (ESI, 40 eV),
m/z (I , %) 254.23 (54) [M] . H NMR spectrum (300 MHz, MeOD-d , δ, ppm, J/Hz): 0.82 (3Í, d, J = 6.5, Ìå-7), 0.89 (3Í,
3
f
3
D
–1
2
+ 1
rel
4
d, J = 6.4, Ìå-9, 10), 0.90 (3Í, d, J = 6.1, Ìå-10, 9), 0.78–1.00 (1Í, m, Í-4α), 1.02–1.32 (3Í, m, Í-2, 3α, 6α), 1.58 (1Í, m,
13
Í-8), 1.82–2.07 (4Í, m, H-3å, 4β, 5, 6β), 4.00 (1Í, br.s, Í-1), 4.85 (2Í, s, NH ). C NMR spectrum (75 MHz, MeOD-d , δ,
2
4
ppm): 21.2 (Ñ-9), 21.6 (Ñ-10), 22.6 (Ñ-7), 27.6 (Ñ-3), 28.3 (Ñ-8), 31.7 (Ñ-5), 36.5 (Ñ-4), 42.7 (Ñ-6), 49.7 (Ñ-2), 50.1 (Ñ-1),
156.3 (Ñ-11), 159.0 (Ñ-14).
ACKNOWLEDGMENT
The work was financially supported in part by the RFBR (Project 16-53-00171 Bel_a) and the UB, RAS, Molecular
Biology Project (No. 18-3-3-17) and used equipment at the Khimiya Common Use Center (CUC), IC, KSC, UB, RAS and the
CUC at the IB, KSC, UB, RAS. Animals from the scientific collection of experimental animals of the IB, KSC, UB, RAS were
used in the work (http://www.ckp-rf.ru/usu/471933/).
887

REFERENCES
1.
2.
N. Siddiqui, W. Ahsan, M. S. Alam, R. Ali, S. Jain, et al., Int. J. Pharm. Sci. Rev. Res., 8 (1), 161 (2011).
D. A. Ibrahim, Eur. J. Med. Chem., 44, 2776 (2009).
3.
4.
5.
I. Ledeti, A. Alexa, V. Bercean, G. Vlase, T. Vlase, L.-M. Suta, and A. Fulias, Int. J. Mol. Sci., 16, 1711 (2015).
S. I. Khan, A. Badshah, M. Aslam, and M. S. Akhtar, JDDMC, 3, 116 (2012).
A. J. K. Atia, Molecules, 14, 2431 (2009).
6.
7.
S. Eswaran, A. V. Adhikari, and N. S. Shetty, Eur. J. Med. Chem., 44, 4637 (2009).
J. Haber, Cas. Lek. Cesk., 140, 596 (2001).
8.
9.
10.
11.
S. Papakonstantinou-Garoufalias, N. Pouli, P. Marakos, and A. Chytyroglou-Ladas, II Farmaco, 57, 973 (2002).
Y. A. Al-Soud, M. N. Al-Dweri, and N. A. Al-Masoudi, II Farmaco, 59, 775 (2004).
B. S. Holla, B. Veerendra, M. Shivananda, and B. Poojary, J. Med. Chem., 38, 759 (2003).
R. Lin, P. J. Connolly, S. Huang, S. K. Wetter, Y. Lu, W. V. Murray, S. L. Emanuel, R. H. Gruninger,
A. R. Fuentes-Pesquera, and C. A. Rugg, J. Med. Chem., 48, 4208 (2005).
12.
T. V. Kochikyan, M. A. Samvelyan, E. V. Arutyunyan, V. S. Arutyunyan, A. A. Avetisyan, M. G. Malakyan,
L. A. Vardevanyan, and S. A. Badzhinyan, Pharm. Chem. J., 44, 525 (2011).
13.
14.
15.
S.-F. Barbuceanu, D. C. Ilies, G. Saramet, V. Uivarosi, C. Draghici, and V. Radulescu, Int. J. Mol. Sci., 15, 10908 (2014).
E. S. Izmest′ev, D. V. Sudarikov, O. G. Shevchenko, S. A. Rubtsova, and A. V. Kutchin, Bioorg. Khim., 41, 90 (2015).
E. V. Buravlev, I. Yu. Chukicheva, O. V. Sukrusheva, O. G. Shevchenko, and A. V. Kutchin, Russ. Chem. Bull., Int. Ed.,
4, 1406 (2015).
16.
17.
18.
19.
Y. V. Gyrdymova, M. Y. Demakova, O. G. Shevchenko, D. V. Sudarikov, L. L. Frolova, S. A. Rubtsova,
and A. V. Kuchin, Chem. Nat. Compd., 53, 895 (2017).
M. Y. Demakova, D. V. Sudarikov, S. A. Rubtsova, L. L. Frolova, A. V. Popov, P. A. Slepukhin, and A. V. Kuchin,
Helv. Chim. Acta, 95, 940 (2012).
M. Y. Demakova, D. V. Sudarikov, S. A. Rubtsova, P. A. Slepukhin, and A. V. Kuchin, Chem. Nat. Compd.,
47, 899 (2012).
Yu. V. Gyrdymova, D. V. Sudarikov, O. G. Shevchenko, S. A. Rubtsova, P. A. Slepukhin, and A. V. Kutchin,
Chem. Biodiversity, 14 (12), e1700296 (2017).
20.
21.
22.
23.
24.
25.
26.
O. V. Dolomanov, L. J. Bourhis, R. J. Gildea, J. A. K. Howard, and H. Puschmann, J. Appl. Crystallogr., 42, 339 (2009).
L. Palatinus and G. Chapuis, J. Appl. Crystallogr., 40, 786 (2007).
G. M. Sheldrick, Acta Crystallogr., Sect. A: Found. Crystallogr., 64, 112 (2008).
H. C. Brown and A. Suzuki, J. Am. Chem. Soc., 89, 1933 (1967).
A. Banach, J. Scianowski, and P. Ozimek, Phosphorus Sulfur Silicon Relat. Elem., 189, 274 (2014).
C. I. Acker, R. Brandao, A. R. Rosario, and C. W. Nogueira, Environ. Toxicol. Pharmacol., 28, 280 (2009).
S. T. Stefanello, A. S. Prestes, T. Ogunmoyole, S. M. Salman, R. S. Schwab, C. R. Brender, L. Dornelles, J. B. T. Rocha,
and F. A. A. Soares, Toxicol. in Vitro, 27, 1433 (2013).
27.
28.
N. A. V. Belle, G. D. Dalmolin, G. Fonini, M. A. Rubim, and J. B. T. Rocha, Brain Res., 1008, 245 (2004).
R. Chawla, R. Arora, R. Kumar, A. Sharma, J. Prasad, S. Singh, R. Sagar, P. Chaudhary, S. Shukla, G. Kaur,
R. K. Sharma, S. C. Puri, K. L. Dhar, G. Handa, V. K. Gupta, and G. N. Qazi, Mol. Cell. Biochem., 273, 193 (2005).
T. Asakawa and S. Matsushita, Lipids, 15, 137 (1980).
J. Takebayashi, J. Chen, and A. Tai, in: Advanced Protocols in Oxidative Stress II, Methods in Molecular Biology,
D. Armstrong (ed.), Humana Press, Totowa, NJ, 2010, Vol. 594, p 287.
29.
30.
31.
J. M. Berg, J. A. F. Kamp, B. H. Lubin, B. Roelofsen, and F. A. Kuypers, Free Radical Biol. Med., 12 (6), 487 (1992).
888