1258
Russ. Chem. Bull., Int. Ed., Vol. 67, No. 7, July, 2018
Kokina et al.
20
[]D +16.0, which was obtained by the distillation of pine
turpentine, sodium nitrite (analytical grade), 35% hydrochloric
acid (reagent grade); 25% aqueous ammonia (analytical grade),
acetyl chloride (high-purity grade), benzaldehyde (Reakhim,
99%), 2-acetylpyridine (Acros, 98%), NaOH (analytical grade),
CuI (Reakhim, analytical grade), diisopropylamine (Acros, 99%),
dimethyl sulfoxide (distilled in vacuo and stored over 4A mo-
lecular sieves), high-purity argon (TU 6-21-12-94), sodium
tartrate dihydrate (Reakhim, analytical grade), and anhydrous
sodium sulfate (analytical grade). The ligand L was purified by
preparative adsorption column chromatography using silica gel
with a mesh size of 50—160 m (IMID); the separation was
controlled by thin-layer chromatography on commercial Sorbfil
plates (silica gel on polyamide film).
added and incubated for 48 h. Then the fluorescent dyes Hoechst
33342 and Propidium iodide were added. The cells were imaged
using an IN Cell Analyzer 2200 system (GE Healthcare, UK) in
an automatic mode for at least four fields per well. The images
were processed with the In Cell Investigator software. The results
are presented as the percentage of live, dead, and apoptotic cells
from three independent experiments standard deviation.
N-{(1S,3S,6R,E)-4-Acetoxyimino-3,7,7-trimethylbicyclo-
[4.1.0]heptan-3-yl}acetamide (C). A solution of acetyl chloride
(10.39 g, 0.1324 mol) in CH2Cl2 (20 mL) was added dropwise
with stirring to a solution of amino oxime C (10.06 g, 0.0552 mol)
and NEt3 (13.37 g, 0.1321 mol) in CH2Cl2 (150 mL) under cool-
ing in an ice bath. The reaction mixture was stirred with cooling
for more 15 min, kept for 10 h at room temperature, and shaken
with 1 M aqueous hydrochloric acid (20 mL). The organic phase
was separated, washed with water until the wash water was neu-
tral, dried with anhydrous Na2SO4, and concentrated in vacuo.
The resulting N,O-diacetyl derivative was used in the synthesis
without purification.
Nitrosyl chloride (NOCl) was prepared from sodium nitrite
and hydrochloric acid and distilled before use. Dimeric crystal-
line nitrosyl chloride of (+)-3-carene was prepared by the reac-
tion with NOCl in dichloromethane as described previously;10
amino oxime B was synthesized from nitroso chloride by the
reaction with excess aqueous ammonia in 95% ethanol.36
(E)-3-Phenyl-1-(pyridin-2-yl)prop-2-en-1-one was prepared by
the condensation of benzaldehyde with 2-acetylpyridine in the
presence of NaOH in water at room temperature as described
previously.37 The complexes were synthesized using CuI (ana-
lytical grade), ethanol (rectified 95%), MeCN, CH2Cl2, and
PriOH (reagent grade).
Microanalyses for C, H, and N were carried out on an
EuroEA 3000 analyzer. The temperature dependence of the
magnetic susceptibility ((T)) of compound 1 was measured on
a Quantum Design MPMSXL SQUID magnetometer in the
temperature range of 2—300 K at a magnetic field strength of
5 kOe. The paramagnetic components of the magnetic suscepti-
bility of the complexes were evaluated taking into account the
diamagnetic contribution estimated from the Pascal constants.
The effective magnetic moment (eff) was calculated by the equa-
tion eff = [3kT/(NAB2)]1/2, where NA, B, and k are Avogadro's
number, the Bohr magneton, and the Boltzmann constant, re-
spectively.
The NMR spectra were recorded on a Bruker DRX-500
spectrometer (1H, 500.13 MHz; 13C, 125.77 MHz) at 27 C for
solutions with a concentration of 25 mg mL–1 in a mixture of
CCl4 and CDCl3 (1 : 1, v/v) using the signals of the solvent CDCl3
(C = 76.90, H = 7.24) as the internal standard. The sign of the
spin-spin coupling constants was not determined. The signal
assignments were made based on 13C NMR spectra recorded in
the J-modulation mode (proton noise-decoupling, the opposite
phases for signals of atoms with odd and even numbers of attached
protons tuned to the constant J = 135 Hz), as well as on two-
dimensional homonuclear 1H—1H correlation spectra, hetero-
nuclear 13C—1H correlation spectra recorded for direct coupling
constants (tuned to the constant J = 135 Hz), and heteronucle-
ar 13C—1H correlation spectra recorded for long-range coupling
constants (tuned to the constant J = 10 Hz). The 13C—1H spin-
spin coupling constants were determined from the 13C NMR
spectra recorded in single resonance mode.
The cell viability was evaluated using an IN Cell Analyzer
2200 system (GE Healthcare, UK), which allows a high-through-
put screening by a method described in the study.35 The Hep2
cell line was cultured in 96-well plates containing IMDM culture
medium, which were placed in a CO2 incubator at 37 C. After
24 h, compounds L and 2 dissolved in DMSO and acetone, re-
spectively (the concentration range 1—50 mol L–1), were
N-{(1aS,3S,7bR)-1,1,3-Trimethyl-7-phenyl-5-(pyridin-2-
yl)-1a,2,3,7b-tetrahydro-1H-cyclopropa[f]quinolin-3-yl}acet-
amide (L). A mixture of N,O-diacetyl derivative C (2.66 g,
10.0 mmol), diisopropylamine (2.02 g, 20.0 mmol), CuI (0.38 g,
2.0 mmol), and (E)-3-phenyl-1-(pyridin-2-yl)prop-2-en-1-one
(2.51 g, 12.0 mmol) in DMSO (50 mL) was kept under an Ar
atmosphere for 48 h at 80 C, cooled to room temperature, and
diluted with an aqueous ammonia solution (50 mL). Then so-
dium tartrate (0.1 g) was added, and the reaction products were
extracted with EtOAc (3Ѕ15 mL). The combined organic extracts
were dried with anhydrous Na2SO4 and concentrated in vacuo.
The resulting dark oily product was chromatographed using the
petroleum ether—EtOAc elution system (10 : 1 1 : 1, v/v).
Ligand L was prepared in a yield of 1.033 g (26% based on N,O-di-
acetyl derivative C) as a fine white powder with m.p. 160—161 C
(from petroleum etherEtOAc) and []27
+120 (c 0.30,
589
CHCl3). High-resolution EI-MS (70 eV). Found: m/z 397.2145
[M]+. C26H27O1N3. Calculated: [M]+ = 397.2149. EI-MS
(70 eV), m/z (Irel (%)): 397 [M]+ (49), 339 [M – CH3CONH]+ (41),
338 [M – CH3CONH2]+ (100), 337 (16), 323 [M – CH3CONH2
– Me]+ (61), 181 (17), 169 (27), 149 (24), 131 (31), 119 (28), 69
(84). IR (КBr), /cm–1: 3288 (N—H), 3050 (Ar—H), 1633
(C=O), 1537 (C=N). 1H NMR (CDCl3—CCl4 1 : 1 v/v), : 0.69
(s, 3 H, H(8)); 0.94 (s, 3 H, H(9)); 1.21 (ddd, 1 H, H(1), J = 9.3 Hz,
J = 8.5 Hz, J = 6.9 Hz); 1.40 (dd, 1 H, pro-S—H(2), J = 15.0 Hz,
J = 6.9 Hz); 1.64 (d, 1 H, H(6), J = 8.5 Hz); 1.90 (s, 3 H, H(12));
1.93 (s, 3 H, H(10)); 3.52 (dd, 1 H, pro-R-H(2), J = 15.0 Hz,
J = 9.3 Hz); 5.37 (s, N—H); 7.23 (ddd, 1 H, H(25), J = 7.4 Hz,
J = 4.8 Hz, J = 1.1 Hz); 7.35—7.44 (m, 5 H, H(17,18,19,20,21));
7.77 (ddd, 1 H, H(24), J = 8.1 Hz, J = 7.4 Hz, J = 1.8 Hz); 8.31
(s, 1 H, H(14)); 8.48 (ddd, 1 H, H(23), J = 8.1 Hz, J = 1.1 Hz,
J = 0.8 Hz); 8.58 (ddd, 1 H, H(26), J = 4.8 Hz, J = 1.8 Hz,
J = 0.8 Hz). 13C NMR (CDCl3 —CCl4, 1 : 1 v/v,), : 16.16 (C(8),
1
1JC,H = 126 Hz); 20.18 (C(1), JC,H = 160 Hz); 23.31 (C(10),
1
1JC,H = 129) Hz; 24.54 (C(6), JC,H = 165 Hz); 24.55 (C(12),
1
1JC,H = 128 Hz); 25.05 (C(7)); 27.25 (C(9), JC,H = 126 Hz);
1
1
30.85 (C(2), JC,H = 132 Hz, JC,H = 128 Hz); 57.27 (C(3));
120.80 (C(14), 1JC,H = 165 Hz, 2,3JC,H < 0.3 Hz); 121.04 (C(23),
1JC,H = 166 Hz, 2,3JC,H = 6.5 Hz); 123.35 (C(25), 1JC,H = 164 Hz,
2,3
J
J
= 8.3 Hz, 2,3JC,H = 6.5 Hz); 127.90 (C(19), 1JC,H = 160 Hz,
C,H
2,3
2,3
= 7.1 Hz,
J
C,H
= 7.1 Hz); 128.00 (C(17), C(21),
= 6.8 Hz); 128.75 (C(18), C(20),
= 6.3 Hz,
C,H
C,H
1JC,H = 160 Hz,
1JC,H = 159 Hz,
J
2,3
2,3
2,3
J
J
= 6.3 Hz); 129.55
C,H
C,H