Synthesis of two new acetanilide derivatives and their effect on the serum antioxidant vitamins (A, E, and C) and the MDA level in rats

Article abstract of DOI:10.1007/s11171-005-0068-7

Acetanilide derivatives, 2,27′-thiobis[N-(4-nitrophenyl)acetamide] and 2,2′-thiobis[N-(4-chlorophenyl)acetamide], were synthesized and characterized. They were shown to cause a considerable oxidative stress in rats.

Full text of DOI:10.1007/s11171-005-0068-7

Russian Journal of Bioorganic Chemistry, Vol. 31, No. 5, 2005, pp. 499–501. Translated from Bioorganicheskaya Khimiya, Vol. 31, No. 5, 2005, pp. 553–556.
Original Russian Text Copyright © 2005 by Karata£, Cansiz, Kara, Karatepe, Koparir.
Synthesis of Two New Acetanilide Derivatives and Their Effect
on the Serum Antioxidant Vitamins (A, E, and C)
and the MDA Level in Rats1
2
F. Karata£*, A. Cansiz*, H. Kara**, M. Karatepe*, and M. Koparir*
* Department of Chemistry, College of Science Department,
F¶rat University, Elaz¶¶, 23169 Turkey
** Elaz¶¶ School of Health Sciences, F¶rat University, Elaz¶¶, 23119 Turkey
Received December 10, 2004; in final form, April 14, 2005
Abstract—Acetanilide derivatives, 2,2'-thiobis[N-(4-nitrophenyl)acetamide] and 2,2'-thiobis[N-(4-chlorophe-
nyl)acetamide], were synthesized and characterized. They were shown to cause a considerable oxidative stress
in rats.
Key words: acetanilide, antioxidant vitamins (A, E, and C), malondialdehyde
INTRODUCTION
To this end, we subcutaneously injected the compounds
to rats and analyzed the serum levels of antioxidant
vitamins (A, E, and C) and MDA, which is the indicator
of lipid peroxidation.
Acetanilide derivatives are reported to exhibit a
number of biological activities, including anesthetic
[1], antipyretic [2, 3], antiinflammatory [4-6] and anti-
3
bacterial [7] effects. In addition, acetaminophen deriv-
ative of acetanilide also is a sedative, a histamin recep-
tor antagonist [8], a painkiller [9–11], and a weak anti-
estrogen [12]. This is the cause that many drugs contain
the moieties of acetanilide and its derivatives in their
molecules.
RESULTS AND DISCUSSION
We chose 2-chloro-N-(4-nitrophenyl)acetamide
(IIa) and 2-chloro-N-(4-chlorophenyl)acetamide, (IIb)
as the starting compounds; they were synthesized by
In this study, we synthesized and characterized two the known procedure [13]. 2,2'-Thiobis[N-(4-nitrophe-
new derivatives of acetanilide, 2,2'-thiobis[N-(4-nitro- nyl)acetamide] (IIIa), and 2,2'-thiobis[N-(4-chlorophe-
phenyl)acetamide] and 2,2'-thiobis[N-(4-chlorophe- nyl)acetamide] (IIIb) were synthesized as described in
nyl)acetamide], and studied their antioxidant activity. [14, 15].
O
O
O
S
ClCH2COCl
K2CO3
Na2S
EtOH
Cl
R–NH
R N
2
NH
R
NH
H
R
(Ia, Ib)
(IIa, IIb)
(IIIa, IIIb)
(a)
R =
;
O2N
Cl
(b) R =
Scheme.
The reaction pathways for the synthesis of (IIa) and the presence of potassium carbonate led to (IIa) and
(IIb) and (IIIa) and (IIIb) are shown in the scheme.
The addition of chloroacetyl chloride to amines (I) in
(IIb). The reaction of (II) with Na2S in ethanol resulted
in aryl-substituted diacetamido sulfides (IIIa) and
(IIIb). The structures of all the compounds were con-
firmed with the help of IR and 1H NMR spectra and the
elemental analysis.
1
The text was submitted by the authors in English.
Corresponding author; e-mail: mkoparir@hotmail.com
2
3
Abbreviations: MDA, malondialdehide.
1068-1620/05/3105-0499 © 2005 Pleiades Publishing, Inc.

500
KARATA‹ et al.
The levels of antioxidant vitamins (A, E and C) and MDA in the serum of control group; nitro group, after the injection of
(IIIa); and chlorine atom, after the injection of (IIIb).*
Group in compound
Antioxidant
Control group
P
chlorine atom in (IIIb) nitro group in (IIIa)
(n = 24)
(n = 24)
Vitamin A, µg/dl
Vitamin E, µg/ml
Vitamin C, µg/ml
MDA, nmol/ml
76.2 + 8.6
9.6 + 1.6
54.1 + 9.2
5.3 + 1.4
8.6 + 1.9
1.51 + 0.3
48.8 + 8.7
5.8 + 1.3
8.9 + 1.7
1.4 + 0.31
<0.001
<0.001
<0.002
<0.001
12.4 + 2.0
0.63 + 0.15
* The number of animals in each group is 24.
The recovery rates were determined to be 98.1% for trophotometer in KBr pellets. NMR spectra were
vitamin A, 99.5% for vitamin E, 96.4% for vitamin C, recorded on a Varian Gemini 200 MHz and Jeol
and 98.2% for MDA. The serum antioxidant vitamins FX-90Q spectrometers and are reported in ppm (δ) rel-
(A, E and C) and MDA levels are given in the table.
ative to tetramethylsilane as an internal standard in deu-
terochloroform. Elemental analyses were done on a
LECO-CHNS-938 instrument. Starting chemicals were
obtained from Merck or Aldrich.
One can see from the table that, being subcutane-
ously injected in rats, the test compounds containing
chlorine (IIIb) and nitro group (IIIa) significantly
decrease the serum levels of vitamins A and E (p <
0.001) and C (p < 0.002) and increase the serum MDA
level (p < 0.001).
The superoxide radicals in organism are removed by
enzymatic dismutation, whereas the antioxidant com-
pounds, such as vitamins A, E, and C, reduce the oxy-
gen radical levels [16] in comparison with control.
General procedure for the synthesis of (IIa) and
(IIb). Chloroacetyl chloride (0.05 mol; in acetone) was
dropwise added to an ice-cooled solution of alkylamine
(0.05 mol) in acetone, and the solution was kept for 2 h
at room temperature. The resulting mixture was poured
into ice-cold water and the crystalline product was col-
lected.
It has been reported that the acetaminophen deriva-
tive of acetanilide damages the liver tissue [17–19] and
increases the nitric oxide formation [6]. It was sug-
gested that high doses of paracetamol decrease the
enzymatic antioxidant level in children and rats [20,
21]. It is also reported that paracetamol catalyzes the
lipid peroxidation [22].
2-Chloro-N-(4-nitrophenyl)acetamide (IIa): yel-
low solid; yield 82%, mp 161–162°ë; IR, ν, cm–1: 1680
1
(C=O), 3277 (NH); H NMR: 7.59–8.66 (5 H, m, Ar
and NH), 4.24 (2 H, s, CH2). Found, %: C 44.68, H
3.11, N 13.00. C8H7ClN2O3. Calc., %: C 44.77, H 3.29,
N 13.05.
2-Chloro-N-(4-chlorophenyl)acetamide
(IIb):
We found that increased MDA levels support the
findings of Kapiotis et al. [22]. Our results showed that
such compounds increase the free radical formation,
which causes an increase in the MDA level as a result
of an enhanced lipid peroxidation. It is also suggested
to use the A, E, and C antioxidant vitamins in order to
make harmless the increased levels of free radicals.
white solid; yield 65%, mp 182–183°ë; IR, ν, cm–1:
1
1670 (C=O), 3263 (NH); H NMR: 8.27 (1 H, br s,
NH), 7.23–7.82 (4 H, m, Ar), 4.21 (2 H, s, CH2). Found,
%: C 44.98, H 3.40, N 6.81. C8H7Cl2NO. Calc., %: C
47.09, H 3.46, N 6.81.
General procedure for the synthesis of (IIIa) and
(IIIb). A solution of sodium sulfide nonahydrate
(0.1 mol) in water (65 ml) was dropwise added to a
stirred boiled solution of (IIa) or (IIb) (0.2 mol) in eth-
anol (200 ml) for 25 min. The addition was slower at
the end to prevent orange coloration.After the reflux for
additional 25 min, the solution was allowed to cool
slowly. Pure crystals were generally obtained directly.
We found no meaningful difference between (IIIa)
and (IIIb) in respect of the vitamins and MDA levels;
both compounds had shown the same effects.
The results concerning the metabolic increase in the
free radical formation by the two derivatives of acet-
anilide allow us to suggest that it is useful to use the
antioxidant vitamins while using medicines that con-
tain acetanilide moieties.
2,2'-Thiobis[N-(4-nitrophenyl)acetamide] (IIIa):
light yellow solid; yield 68%; mp 220–221°C; IR, ν,
cm–1: 1679 (C=O), 3280 (NH); 1H NMR: 10.27 (2 H, br
s, NH), 7.45–8.54 (8 H, m, Ar), 3.57 (4 H, s, CH2S).
Found, %: C 49.29, H 3.59, N 14.21, S 8.31.
C16H14N4O6S. Calc., %: C 49.23, H 3.61, N 14.35, S,
EXPERIMENTAL
Melting points were determined on a Thomas
Hovver melting point apparatus and were not corrected.
A differential scanning calorimeter (DSC) was used. IR
spectra were measured on a Mattson 1000 FTIR spec- 8.21.
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 31 No. 5 2005

SYNTHESIS OF TWO NEW ACETANILIDE DERIVATIVES AND THEIR EFFECT
501
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2,2'-Thiobis[N-(4-chlorophenyl)acetamide] (IIIb):
shine white solid; yield 74%; mp 271–272°ë; IR, ν,
cm–1: 1671 (C=O), 3268 (NH); 1H NMR: 9.98 (2 H, br
s, NH), 7.40–7.70 (8 H, m, Ar), 3.54 (4 H, s, CH2S).
Found, %: C 52.01, H 3.79, N 7.51, S 8.59.
C16H14Cl2N2O2S. Calc., %: C 52.04, H 3.82, N 7.59, S
8.68.
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