Synthesis and Biological Activity of Aliphatic and Aromatic Sulfonic Acid Azolides
589
TABLE 1. Yields and Physicochemical Characteristics of Compounds I – IV
Com-
pound
Empirical
formula
IR spectrum:
Yield, %
85
1H NMR chemical shift: d, ppm
nmax, cm – 1
I
C7H7N3O2S
C8H8N2O2S
C8H7N3O2S
1040 (SO2)
1380 (SN)
7.91 – 7.88 (m, 2H), 7.43 – 7.39 (m, 2H), 2.57 (s, 3H, CH3)
II
1050 (SO2)
1350 (SN)
87
85
8.55 (s, 1H), 7.86 – 7.81 (m, 2H), 7.48 – 7.43 (m, 2H), 3.70 (s, 3H, CH3)
III
3160,
10.00 (s, 2H, H-triaz.), 9.16 (s, 1H, H-2¢-phenyl), 7.51 (m, 1H,
H-3¢-phenyl), 7.49 (m, 1H, H-3¢-phenyl), 7.11 (m, 1H, H-4¢-phenyl), 7.10
(s, 1H, H-5¢-phenyl)
3100 (C-Htriaz),
3070 (C-Hphenyl
)
IV
C9H8N2O2S
3150,
85
7.58 (m, 1H, H-2-imidaz.), 8.84 (s, 1H, H-4-imidaz.), 7.39 (s, 1H,
H-2¢-phenyl), 7.29 (m, 1H, H-3¢-phenyl), 7.14 (m, 1H, H-4¢-phenyl), 7.12
(m, 1H, H-5¢-phenyl), 7.11 (m, 1H, H-5¢-phenyl)
3120 (C–Himidaz.), 3070
(C–Hphenyl
)
Cytotoxicity of the synthesized com-
TABLE 2. Cytotoxic and Cytoprotector Effect of Aliphatic and Aromatic Sulfonic Acid
pounds was studied using an endothelial
cell culture grown in modified Petri dishes
[3]. The funic endothelium was taken from
an Rh-positive fetus with blood group 0(I)
obtained from physiologically pregnant fe-
males. The cell dissociation was provided
by a combined enzymatic-mechanical tech-
nique. The cytotoxicity was evaluated by
counting viable and lost cells after a 96-h in-
cubation of the cell cultures with test com-
pounds and ethidium bromide and acridine
orange staining [3].
The immunomodulant properties were
estimated using an enzymatic test with pe-
ripheral blood monocytes (PBMs), by de-
termining the activity of pair glycolytic en-
zymes a-glycerophosphatedehydrogenase
(a-GPDG) and succinatedehydrogenase
(SDG). The test was performed with the
Azolides with Respect to Endothelial Cell Culture
Percentage
cell survival
in culture
Compound
(n, number
of tests)
Percentage cell
loss in culture
with AEABs
Percentage
cell survival
Percentage
cell loss
with AEABs
I (n = 19)
96.2 ± 3.4
94.3 ± 2.9
95.3 ± 4.1
90.5 ± 6.8
21.3 ± 12.9
3.8 ± 1.6
5.7 ± 2.0
4.7 ± 1.9
9.5 ± 2.3
78.7 ± 14.6
53.4 ± 13.2*
20.2 ± 9.6
64.3 ± 11.5*
39.6 ± 8.7
–
46.6 ± 10.2*
79.8 ± 14.3
35.7 ± 9.9*
60.4 ± 12.3
–
II (n = 21)
III (n = 24)
IV (n = 18)
Control 1 (with AEABs)
(n = 17)
Control 2 (without
drugs, with AEABs)
(n = 14)
–
–
92.1 ± 3.6
7.9 ± 2.1
Note. P < 0.05 relative to control 1.
blood of (i) females with a physiological course of preg-
nancy (23 cases) possessing a relative (physiological) immu-
nodeficiency and (ii) females with hestosis (32 cases) featur-
ing a-GPDG and SDG activity [4]; a group of 26 healthy
nonpregnant females of the corresponding age served as the
control group. The a-GPDG and SDG activity in PBMs was
determined by microscopic examination of peripheral blood
smears after conventional cytochemical processing [5]. The
PBM were isolated by the ficoll – hypaque gradient method
[3]. The immunomodulant effect of the synthesized com-
pounds was evaluated 72 h after introduction into the cul-
tures of monocytes taken from females of both test and con-
trol groups.
The index of cytotoxicity (IC) of the sera containing
anti-endothelial antibodies (AEABs) was evaluated accord-
ing to the NIAID standard (National Institute of Allergies
and Infectious Diseases, USA) using the monocyte cross test
based on the antigen concordance between endotheliocytes
and monocytes [6]. For this analysis, monocytes used as the
test cells were isolated from funic blood of healthy Rh-posi-
tive fetus with blood group 0(I). The cells were incubated for
30 min with serum, and then for 2 h with the complement.
The reacted cells were fixed with formaldehyde and stained
with eosin. The IC of AEABs was rated against the following
scale depending on the content of damaged cells: negative
result, below 10%; weak positive result, 10 – 20%; positive
result, 20 – 50%; sharply positive result, 50 – 100%.
RESULTS AND DISCUSSION
According to the test results, the mutagenicity of com-
pounds I – IV is characterized by the following values.
Methanesulfonic acid benzotriazolide (I) induced 1.18% re-
cessive lethal mutations in X-chromosomes of Drosophila
menogaster males, while benzenesulfonic acid 1,2,4-triazo-
lide benzimidazolide (III) induced 1.97% such lethals.
Benzenesulfonic acid imidazolide (IV) induced 6.95% domi-
nant lethal mutations, while methanesulfonic acid
benzimidazolide (II) induced 21% such mutations in both