Synthesis and evaluation of anti-inflammatory and antitussive activity of hydantion derivatives

Article abstract of DOI:10.2174/157018012800673092

1-Methylhydantion is a compound, which was isolated from Oviductus Ranae for the first time. In our study, we found that it showed good antitussive and anti-inflammatory activity. It is also the first report which illustrates the antitussive activity of hydantion derivative. A series of hydantion derivatives were synthesized and evaluated for their anti-inflammatory and antitussive activity in vivo. The pharmacological tests showed that compounds 7a, 7c and 7d have good anti-inflammatory and antitussive activity compared to Ibuprofen and codeine. Compound 7a in particular showed two-fold stronger anti-inflammatory activity than Ibuprofen.

Full text of DOI:10.2174/157018012800673092

638
Letters in Drug Design & Discovery, 2012, 9, 638-642
Synthesis and Evaluation of Anti-Inflammatory and Antitussive Activity of
Hydantion Derivatives
Haibin Lu*,1, Dejuan Kong1, Bin Wu1, Shihan Wang2 and Yongsheng Wang*,1
2
1College of Pharmacy, Jilin University, Changchun 130021, P. R. China; Liaoning University of Traditional Chinese
Medicine, Shenyang 116600, P. R. China
Received February 04, 2012: Revised March 11, 2012: Accepted April 13, 2012
Abstract: 1-Methylhydantion is a compound, which was isolated from Oviductus Ranae for the first time. In our study,
we found that it showed good antitussive and anti-inflammatory activity. It is also the first report which illustrates the
antitussive activity of hydantion derivative. A series of hydantion derivatives were synthesized and evaluated for their
anti-inflammatory and antitussive activity in vivo. The pharmacological tests showed that compounds 7a, 7c and 7d have
good anti-inflammatory and antitussive activity compared to Ibuprofen and codeine. Compound 7a in particular showed
two-fold stronger anti-inflammatory activity than Ibuprofen.
Keywords: Anti-inflammatory activity, Antitussive activity, Biological activity, Hydantoin, Ibuprofen, Synthesis.
INTRODUCTION
molecule. A series of hydantion derivatives were synthesized
and evaluated for their anti-inflammatory and antitussive
activity in vivo. The pharmacological tests showed that
compounds 7a, 7c and 7d have good anti-inflammatory and
antitussive activity compared to Ibuprofen and codeine.
Compound 7a even showed two-fold stronger anti-
inflammatory activity than Ibuprofen.
Hydantoin (1, 3-imidazolidinedione) was discovered in
1861 by Baeyer, who isolated it as one of the reduction
products of allantoin during the study of uric acid.
Hydantoin derivatives display diverse and interesting
pharmacological
properties.
Such
as
phenytoin,
mephenytoin, norantoin, and some 5-substituted hydantoins
are well-known anticonvulsive drugs [1, 2, 3]. Hydantoins
also exhibit antidepressant, antiviral, antitumoral and
antithrombotic activities, as well as inhibitory activity
against some enzymes such as human aldose reductase and
human leucocyte elastase [4].
METHODS
Evaluation of Anti-Inflammatory Activity [8]
Anti-inflammatory activity of these hydantion derivatives
was evaluated using xylene-induced mouse ear edema
model. Kunming male mice (20-22g) were divided into ten
groups randomly (n=10 for each group). One group was kept
as control and the others were given test compounds
including one group given Ibuprofen as standard. 1 hour
after the last oral treatment, 0.02mL xylene was applied to
the right ear of each mouse. Mice were killed by cervical
dislocation 1 hour after xylene application. Ear disk of 8.0
mm in diameter was punched out and weighed. The
difference in weight of the ears from each group was used to
analyze the anti-inflammatory function.
Oviductus Ranae is the dry oviductus of the tely-Rana
temporaria chensinensis David, which has many
pharmacological actions, such as enhancement of immune
and stress performance, antioxidant and anti-aging, blood fat
regulation, the effect of growth and sexual function, anti-
hypoxia and anti-anxiety, anti-fatigue, antitussive
expectoration and ataxia regulation [5]. 1-Methylhydantion,
a hydantion derivative, for the first time has been isolated
from Oviductus Ranae [6]. In our study, it shows good
antitussive activity and anti-inflammatory activity. It is the
first report which shows that hydantion derivative has
antitussive activity.
In this paper, we report our efforts towards the synthesis
and investigation of the antitussive activity of a series of
novel hydantoin derivatives. We have also optimized the
compound to enhance the anti-inflammatory activity of the
hydantion derivatives. This allows the compound to possess
both good antitussive activity and good anti-inflammatory
Evaluation of Antitussive Activity [9]
The antitussive activity was evaluated in mouse after
cough induction by ammonia aerosol stimulation and the
number of cough was detected in 2 min. Kunming male mice
(20-22g) were divided into ten groups randomly (n=10 for
each group). One group was kept as control and the others
were given test compounds including one group given
codeine as standard. 1 hour after the last oral treatment, mice
cough was induced by ammonia aerosol stimulation. The
antitussive effects of the target compounds on mice were
evaluated by the frequency of cough caused by ammonia and
the latent period of cough after administration.
activity. Ibuprofen,
a
common non-steroidal anti-
inflammatory drug (NSAIDs) is used primarily for fever,
pain, dysmenorrhea and inflammatory diseases [7]. Here we
combined the structure of Ibuprofen and hydantoin into one
*Address correspondence to these authors at the College of Pharmacy, Jilin
University; Ltd., No. 1266 Fujin Road, changchun, 130021, People’s
Republic of China; Tel: +86-0431-85619121; Fax: +86-0431-85619121;
E-mails: haibin1025@yahoo.cn and mikewangwys@163.com
1875-628X/12 $58.00+.00
© 2012 Bentham Science Publishers

Hydantion Derivatives Synthesis and Activity Evaluation
Letters in Drug Design & Discovery, 2012, Vol. 9, No. 6 639
O
R1
O
O
N
O
HCl , KCNO
H
HCl
HN
N
R1
O
NH2
N
R1
O
O
1a-b
2a-b
3a-b
O
NC
R3
OH
R2
(NH4)2CO3
Na2S2O5, KCN
O
HN
NH
R3
R2
R3
O
R2
5a-b
4a-b
3c-d
R3
N
O
O
R2
R1
SOCl2, Pyridine
OH
HN
O
N
+
N
R1
O
R2
O
O
R3
3a-d
6
7a-d
7a:
7b:
R1= -CH3 ,R2=H , R3=H
R1=-CH2Ph ,R2=H , R3=H
3a: R1= -CH3
3b: R1= -CH2Ph
3c: R2 = H, R3 = H
3d: R2= -CH3, R3 = -CH3
7c:
7d:
R1=H ,R2=H , R3=H
R1= -CH3 ,R2=-CH3 , R3=-CH3
Scheme 1. Synthesis of hydantion derivatives.
RESULTS AND DISCUSSION
Synthesis
Biological Activity
The anti-inflammatory activity and antitussive activity of
hydantion derivatives were summarized in Tables 1 and 2.
The results showed that compounds 7a, 7c and 7d have both
good anti-inflammatory and antitussive activity. It should be
noted that compound 7a has two-fold stronger anti-
inflammatory activity than Ibuprofen. Compounds 3a, 3c
and 3d showed good antitussive activity, but anti-
inflammatory activity was weak. Compounds 3b and 7b that
substituted benzyl at N1 position of hydantoin have nearly no
activity.
A general synthetic route for the hydantion derivatives
used in this study is shown in Scheme 1. In order to reduce
the toxicity dangerous, compounds 3a-d were synthesized by
one pot method. Hydantoins 3a-b were prepared by acid-
catalyzed cyclization of N-substituted glycine ethylesters 1a-
b [10]. N-substituted glycine ethylesters 1a-b were first
reacted with potassium cyanate to obtain 2a-b, Compounds
2a-b were then cyclized under acidic condition to get 3a-b.
5-Substituted hydantoins 3c-d were synthesized through
Bucherer-Berg method [11, 12]. Acetone or formaldehyde
were first reacted with potassium cyanide to get cyanohydrin
derivatives 5a-b, Compounds 5a-b were then treated with
ammonium carbonate to obtain 3c-d. Ibuprofen was first
treated with SOCl2 to obtain the acyl chloride, and then
reacted with hydantoins 3a-d in pyridine to give the target
compounds 7a-d. The structures of compounds were
confirmed by MS, 1H NMR, 13C NMR and Elemental
analysis.
These SAR studies revealed that the substitution of
methyl or hydrogen at the N1 position of hydantoin had
effective influence on antitussive activity. Especially, N1
substituted 1-methyl hydantoin 7a had better anti-
inflammatory activity than Ibuprofen. However, with benzyl
group at N1 position only showed weak anti-inflammatory
and antitussive activity. Because compound 7b didn’t show
any anti-inflammatory activity, so the mechanism of anti-
inflammatory action may not be as that the hydantion
derivatives first hydrolyze to Ibuprofen, then Ibuprofen plays

640 Letters in Drug Design & Discovery, 2012, Vol. 9, No. 6
Lu et al.
Table 1. Anti-Inflammatory Activity of the Target Compounds
Compound
Dose(mg/kg)
Ear swelling degree a (mg)
Inhibition rate b (%)
control
5%CMC
100
12.36±0.52*
8.89±0.33**
10.58±0.32*
12.15±0.42*
10.60±0.44*
10.50±0.58*
5.61±0.42**
11.38±0.46**
9.33±0.51*
Ibuprofen
28.07
14.35
1.70
3a
3b
3c
3d
7a
7b
7c
7d
100
100
100
14.19
15.08
54.61
7.93
100
100
100
100
24.51
25.00
100
9.27±0.40**
a Ear swelling degree was calculated as (the weight of right ear)-(the weight of left ear)
b Inhibition rate was calculated as（1-the ear swelling degree of the compound/the ear swelling degree of the control）×100%
Standard: Ibuprofen, each value is the mean ± SEM for ten mice,* P < 0.05;** P < 0.01 compared with control.
Data analyzed by one-way ANOVA followed by Student’s t-test
Table 2. Antitussive Effect of the Target Compounds
Compound
Dose(mg/kg)
Latency (s)
Frequency of cough
control
codeine
3a
5%CMC
30
27.3±5.8*
53±6.2*
42±3.1*
17±2.7**
16.5±2.5*
45.5±3.5*
17.5±4.5*
17.9±3.8*
15.5±3.5**
29.2±3.3*
18.3±3.6**
18.6±2.9**
100
39.5±9.2*
29.5±6.29*
36.3±5.3*
33.5±4.6*
40±4.8*
3b
100
3c
100
3d
100
7a
100
7b
100
33.3±7.1**
77±6.7*
7c
100
7d
100
38.3±5.2*
Standard: codeine, each value is the mean ± SEM for ten mice,* P < 0.05;** P < 0.01 compared with control.
Data analyzed by one-way ANOVA followed by Student’s t-test.
the anti-inflammatory action. Further extensive studies need
to be carried out to determine their mechanism of anti-
inflammatory action.
0.028 mol) in 3.24 mL of water was added dropwise. The
mixture was stirred at room temperature for 20 h and then
extracted with CH2Cl2. The organic layer was dried under
reduced pressure to get the crude N-substituted N-
carbethoxymethylureas 2. 12.50 mL 25% HCl was then
added into the mixture and heated under reflux for 4 h and
then cooled in ice. The precipitate was collected and
recrystallized with hot water to get pure product 3a-b as
white solid.
SYNTHESIS EXPERIMENTAL SECTION
All reagents were of commercial quality and were used
as received. Solvents were dried and purified by using
standard techniques. Reactions were monitored by TLC. The
elemental analysis was performed on a Carlo Erba 1106
1-Methylimidazolidine-2, 4-Dione (3a)
1
analyzer. The H NMR and 13C NMR spectra were recorded
1
Yield 62%; H-NMR（CDCl3, 400 MHz） δ: 2.98 (s,
on a UNITY-400 NMR spectrometer with TMS as the
internal standard. The MS spectra were taken on an LCQ
electrospray mass spectrometer.
3H, CH3), 3.93 (s, 2H, CH2), 8.74 (s, 1H, NH); 13C-
NMR（CDCl3, 100 MHz） δ: 170.22, 156.66, 52.93, 29.37.
1-Benzylimidazolidine-2, 4-Dione (3b)
Yield 68%; 1H-NMR（CDCl3, 400 MHz） δ:3.77 (s, 2H,
CH2), 4.52 (s, 2H, CH2), 7.24-7.29 (m, 2H, 2×ArH), 7.31-
7.37 (m, 3H, 3×ArH), 9.27 (s, 1H, NH); 13C-NMR（CDCl3,
General Procedure for the Compounds 3a-b
N-substituted glycine ethylester 1 (0.019 mol) was added
into conc. HCl (2.4 mL, 0.028 mol). With continued stirring
at room temperature, a solution of potassium cyanate (2.3 g,

Hydantion Derivatives Synthesis and Activity Evaluation
Letters in Drug Design & Discovery, 2012, Vol. 9, No. 6 641
100 MHz） δ: 171.44, 156.96, 136.52, 128.53, 127.42,
127.42, 127.33, 127.33, 50.38, 45.26.
379.4; Anal. Calcd. for C23H26N2O3: C 72.99, H 6.92, N
7.40; Found C 72.96, H 6.91, N 7.41.
3-(2-(4-Isobutylphenyl)propanoyl)Imidazolidine-2,4-Dione
(7c)
General Procedure for the Compounds 3c-d
White solid, yield 75%; 1H-NMR（CDCl3, 400 MHz）δ:
0.87 (d, J = 6.64 Hz, 6H, 2×CH3), 1.49 (d, J = 6.96 Hz, 3H,
CH3), 1.83-1.85 (m, 1H, CH), 2.42 (d, J = 7.16 Hz, 2H,
CH2), 3.86 (d, J = 4.76 Hz, 2H, CH2), 4.81 (q, J = 6.96 Hz,
1H, CH), 5.94 (s, 1H, NH), 7.07 (d, J = 8.12 Hz, 2H,
2×ArH), 7.15 (d, J = 8.12 Hz, 2H, 2×ArH); 13C-
NMR（CDCl3, 100 MHz）δ：172.72, 168.52, 154.86,
141.07, 136.43, 129.66, 129.66, 127.61, 127.61, 46.96,
45.71, 44.99, 30.08, 22.35, 22.35, 18.44; ESI-MS (M++H),
m/z: 289.3; Anal. Calcd. for C16H20N2O3 : C 66.65, H 6.99, N
9.72; Found C 66.66, H 6.99, N 9.74.
Sodium metabisulphite (14.6 g, 0.077 mol), compound 4
(0.077 mol) and potassium cyanide (5.0 g, 0.077 mol) was
dissolved in 20 mL water, stirred at room temperature for 0.5
h to get cyanohydrin derivatives 5. Ammonium carbonate
(14.8 g, 0.154 mol) was then added. The mixture was heated
to 50℃ for half an hour, and the temperature was raised to
90℃ and heated for another 3 h. The solvent was evaporated
under reduced pressure. The crude product was recrystallized
with hot water to get pure product 3c-d as white solid.
Imidazolidine-2, 4-Dione (3c)
Yield 42%; 1H-NMR（DMSO-d6, 400 MHz） δ: 3.84 (s,
2H, CH2), 7.67 (s, 1H, NH), 10.59 (s, 1H, NH); 13C-
NMR（DMSO-d6, 100 MHz） δ: 174.26, 158.74, 47.56.
3-(2-(4-Isobutylphenyl)propanoyl)-5,5-
Dimethylimidazolidine-2,4-Dione (7d)
White solid, yield 85%; 1H-NMR （CDCl3, 400
MHz）δ：0.85 (d, J = 6.6 Hz, 6H, 2×CH3), 1.23 (d, J = 11.2
Hz, 6H, 2×CH3), 1.48 (d, J = 6.96 Hz, 3H, CH3), 1.80-1.82
(m, 1H, CH), 2.42 (d, J = 7.16 Hz, 2H, CH2), 4.75 (q, J =
6.96 Hz, 1H, CH), 6.08 (s, 1H, NH), 7.06 (d, J = 8.0 Hz, 2H,
5, 5-Dimethylimidazolidine-2, 4-Dione (3d)
Yield 48%; 1H-NMR（DMSO-d6, 400 MHz） δ: 1.44 (s,
6H, 2×CH3), 7.82 (s, 1H, NH), 10.68 (s, 1H, NH); 13C-
NMR（DMSO-d6, 100 MHz） δ: 181.78, 158.49, 61.35,
24.26.
2×ArH), 7.11 (d,
J =
8.0 Hz, 2H, 2×ArH); 13C-
NMR（CDCl3, 100 MHz）δ：174.39, 172.89, 152.60,
141.01, 136.75, 129.58, 129.58, 127.62, 127.62, 58.02,
47.17, 44.93, 30.1, 24.92, 24.92, 22.24, 22.24, 17.93; ESI-
MS (M++H), m/z: 317.4, Anal. Calcd. for C18H24N2O3 : C
68.33, H 7.65, N 8.85; Found C 68.34, H 7.64, N 8.86.
General Procedure for the Compounds 7a-d
Ibuprofen (0.5 g, 0.002 mol), SOCl2(0.48 g, 0.004 mol)
added into CH2Cl2 10 ml, stirred at reflux for 2 h, then added
into compound 3 (0.002 mol) were dissolved in pyridine 10
mL, reacted at room temperature overnight. By evaporating
the solvent, compound 7 was obtained through flash
chromatography (methanol / chloroform 1/20).
CONCLUSION
In summary, a series of novel hydantion derivatives were
synthesized and evaluated for their anti-inflammatory and
antitussive activity. These SAR studies revealed that the
substitution of methyl and hydrogen at the N1 position of
hydantoin had pronounced influence on antitussive activity.
It's noteworthy to mention that compound 7a showed two-
fold stronger anti-inflammatory activity than Ibuprofen.
Further extensive studies are required to be carried out to
determine their mechanism of action.
3-(2-(4-Isobutylphenyl)Propanoyl)-1-Methylimidazolidine-
2,4-Dione (7a)
White solid, yield 82%; 1H-NMR（CDCl3, 400
MHz）δ：0.89 (d, J = 6.8 Hz, 6H, 2×CH3), 1.49 (d, J = 6.8
Hz, 3H, CH3), 1.83-1.85 (m, 1H, CH), 2.42 (d, J = 7.2 Hz,
2H, CH2), 2.93 (s, 3H, CH3), 3.77 (s, 2H, CH2), 4.85 (q, J =
6.8 Hz, 1H, CH), 7.07 (d, J = 8.0 Hz, 2H, 2×ArH), 7.16 (d, J
= 8.0 Hz, 2H, 2×ArH); 13C-NMR（CDCl3, 100 MHz）δ：
172.58, 166.93, 152.86, 140.88, 136.68, 129.57, 129.57,
127.64, 127.64, 50.85, 46.62, 44.97, 30.06, 29.52, 22.32,
22.32, 18.57; ESI-MS (M++H), m/z: 303.3; Anal. Calcd. for
C17H22N2O3: C 67.53, H 7.33, N 9.26; Found C 67.55, H
7.32, N 9.25.
CONFLICT OF INTEREST
Declared none.
ACKNOWLEDGEMENTS
1-Benzyl-3-(2-(4-isobutylphenyl)Propanoyl)Imidazolidine-
2,4-Dione (7b)
This work was supported by the Natural Medicinal
Chemistry Department of the College of Pharmacy Jilin
University.
Oil， yield 90%; 1H-NMR（CDCl3, 400 MHz）δ：0.89
(d, J = 6.56 Hz, 6H, 2×CH3), 1.50 (d, J = 6.92 Hz, 3H, CH3),
1.84-1.86 (m, 1H, CH), 2.44 (d, J = 7.16 Hz, 2H, CH2), 3.60
(dd, J1 = 17.72 Hz, J2 = 25.0 Hz, 2H, CH2), 4.46 (dd, J1 =
14.96 Hz, J2 = 6.4 Hz, 2H, CH2), 4.87 (q, J = 6.96 Hz, 1H,
CH), 7.11-7.18 (m, 6H, 6×ArH), 7.32 (m, 3H, 3×ArH); 13C-
NMR（CDCl3, 100 MHz）δ: 172.60, 166.88, 152.84,
140.93, 136.69, 134.42, 129.60, 129.60, 129.02, 129.02,
128.33, 128.00, 128.00, 127.68, 127.68, 48.24, 46.83, 46.44,
45.00, 30.10, 22.34, 22.34, 18.39; ESI-MS (M++H), m/z:
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