Synthesis and antimicrobial activity of some new substituted pyrido[3′,2′:4,5]thieno[3,2-d]-pyrimidinone derivatives

Article abstract of DOI:10.1134/S1068162014030042

A series of new 3-substituted-7-(2-chloro-6-ethoxypyridin-4-yl)-9-(2,4- dichlorophenyl)-2-methylpyrido[3′,2′:4,5]thieno[3,2-d] pyrimidin-4(3H)-one derivatives were synthesized as antimicrobial agents using 7-(2-chloro-6-ethoxypyridin-4-yl)-9-(2,4-dichlorophenyl)-2-methyl-4H- pyrido[3′,2′:4,5]thieno[3,2-d]-[1,3]oxazin-4-one as a starting compound. Its condensation with substituted aniline derivatives or phenyl hydrazine gave the corresponding N-substituted derivatives. Treatment of the starting compound with hydrazine hydrate afforded the corresponding N-amino derivative, which was reacted with substituted phenylisocyanate and phenylisothiocyanate derivatives to give the corresponding semicarbazides and thiosemicarbazide derivatives. All the newly synthesized compounds were evaluated for their antimicrobial activities in comparison to streptomycin and fusidic acid as positive controls. The structure assignments of the new compounds are based on chemical and spectroscopic evidence.

Full text of DOI:10.1134/S1068162014030042

ISSN 1068ꢀ1620, Russian Journal of Bioorganic Chemistry, 2014, Vol. 40, No. 3, pp. 308–313. © Pleiades Publishing, Ltd., 2014.
Synthesis and Antimicrobial Activity of Some New Substituted
Pyrido[3',2':4,5]thieno[3,2ꢀd
]ꢀPyrimidinone Derivatives1
Ahmed A. Fayeda, b, 2, Abd ElꢀGalil E. Amrb, c, Mohamed A. AlꢀOmarc, and Elsayed E. Mostafad
a Respiratory Department, Applied Organic & Biochemistry Division, College of Medical Rehabilitation Sciences,
Taibah University, AlꢀMadinah AlꢀMunawara, 3893 Saudi Arabia
b Applied Organic Chemistry Department, National Research Center, Cairo, Dokki, Egypt
c Pharmaceutical Chemistry Department, Drug Exploration & Development Chair (DEDC),
College of Pharmacy, King Saud University, Riyadh, 11451 Saudi Arabia
d Department of Microbial Chemistry, National Research Center, Cairo, Egypt
Recevied November 21, 2013; in final form, December 11, 2013
Abstract
methylpyrido[3',2':4,5]thieno[3,2ꢀ
using 7ꢀ(2ꢀchloroꢀ6ꢀethoxypyridinꢀ4ꢀyl)ꢀ9ꢀ(2,4ꢀdichlorophenyl)ꢀ2ꢀmethylꢀ4
[1,3]oxazinꢀ4ꢀone as a starting compound. Its condensation with substituted aniline derivatives or phenyl
hydrazine gave the corresponding ꢀsubstituted derivatives. Treatment of the starting compound with hydraꢀ
zine hydrate afforded the corresponding ꢀamino derivative, which was reacted with substituted phenylisoꢀ
—
A series of new 3ꢀsubstitutedꢀ7ꢀ(2ꢀchloroꢀ6ꢀethoxypyridinꢀ4ꢀyl)ꢀ9ꢀ(2,4ꢀdichlorophenyl)ꢀ2ꢀ
]pyrimidinꢀ4(3 )ꢀone derivatives were synthesized as antimicrobial agents
ꢀpyrido[3',2':4,5]thieno[3,2ꢀ ]ꢀ
d
H
H
d
N
N
cyanate and phenylisothiocyanate derivatives to give the corresponding semicarbazides and thiosemicarbaꢀ
zide derivatives. All the newly synthesized compounds were evaluated for their antimicrobial activities in
comparison to streptomycin and fusidic acid as positive controls. The structure assignments of the new comꢀ
pounds are based on chemical and spectroscopic evidence.
Keywords: oxazinone, thienopyrimidine, semiꢀ and thiosemicarbazides, antimicrobial agents
DOI: 10.1134/S1068162014030042
2 1
INTRODUCTION
RESULTS AND DISCUSSION
In our previous work, some of fused heterocyclic
In the present work, 2ꢀchloroꢀ6ꢀethoxyꢀ4ꢀ[(2ꢀmeꢀ
thylꢀ4ꢀoxoꢀ9ꢀ(2,6ꢀdichlorophenyl)pyrido[3',2':4,5]ꢀ
thieno[3,2ꢀd]oxazinꢀ7ꢀyl]pyridines (II) was used as
starting material and it was prepared from the correꢀ
sponding 1ꢀ(2ꢀchloroꢀ6ꢀethoxypyridinꢀ4ꢀyl)ꢀ3ꢀ(2,6ꢀ
dichlorophenyl)propꢀ2ꢀenꢀ1ꢀone (I) according to the
reported methods [1] (Scheme 1).
derivatives were studied as antimicrobial [1],
5 ꢀ
α
reductase inhibitors, antiviral, antiꢀtumor, antiꢀparꢀ
kinsonism, hypoglycemic, antiꢀmicrobial, antiꢀ
inflammatory, analgesic, and antipyretic agents [2–9].
In addition, biological and antianalgesic activities of
many heterocyclic compounds containing a sulfur
atom have been reviewed [10–12]. On the other hand,
thienopyrimidine and thioxopyrimidine derivatives
have promising biological [13, 14] and anticancer
activities [15, 16]. Recently, some new substituted
thienopyrimidine derivatives were synthesized [17]
and evaluated as antiꢀinflammatory [18], antiproliferꢀ
ative [19], and analgesic [20] agents. In view of these
observations and as continuation of our previous
works in heterocyclic chemistry, we have herein synꢀ
thesized some new thienopyrimidine derivatives and
tested their antiꢀmicrobial activities.
Treatment of oxazinone derivatives (IIa, b) with substiꢀ
tuted aniline derivatives or phenylhydrazine in glacial
acetic acid under reflux condition afforded the correꢀ
sponding 7ꢀ(2ꢀchloroꢀ6ꢀethoxypyridinꢀ4ꢀyl)ꢀ9ꢀ(2,6ꢀ
dichlorophenyl)ꢀ2ꢀmethylꢀ3ꢀ[
nyl)pyrido[3',2':4,5]thieno[3,2ꢀ ]pyrimidineꢀ4(3
IIIa ) and 7ꢀ(2ꢀchloroꢀ6ꢀethoxypyridinꢀ4ꢀyl)ꢀ9ꢀ(2,6ꢀ
dichlorophenyl)ꢀ3ꢀphenylꢀ2ꢀmethylpyrido[3',2':4,5]ꢀ
thieno[3,2ꢀ ]pyrimidineꢀ4(3 )ꢀone (IV), respecꢀ
N
ꢀ(substituted pheꢀ
d
H)ꢀone
(
–e
d
H
tively. In addition, reacting of (II) with hydrazine hyꢀ
drate in refluxing ethanol afforded the corresponding
N
reacted with substituted phenylisothiocyanate to give the
ꢀaminothienopyrimidine derivative (
V), which was
1
corresponding thiosemicarbazide derivatives (VIa–e), reꢀ
spectively (Scheme 2).
The article is published in the original.
Corresponding author: eꢀmail: dr_ahmedfayed14@yahoo.com.
2
308

SYNTHESIS AND ANTIMICROBIAL ACTIVITY
Cl
309
CH3
O
Cl
Cl
Cl
Cl
CHO
N
Cl
OEt
O
N
Cl
OEt
I
Cl
S
N
O
CH3
Cl
O
N
N
OEt
II
Scheme 1. Synthetic pathway for starting material (II).
Antimicrobial activities of the newly synthesized compounds (III
–VI)
Fungi Yeast
Bacteria
Comp. no.
Gramꢀpositive
Gramꢀnegative
A. n.
Pen. sp.
C. a.
R. i.
B. s.
S. l.
Str. sp.
E. c.
P. sp.
(
(
(
(
(
(
(
(
(
(
(
(
(
(
IIIa
)
)
)
10
7
13
5
10
8
11
9
20
12
13
11
13
20
13
11
23
13
23
11
23
20
22
–
21
13
12
10
12
21
12
10
24
12
24
12
22
21
21
–
21
6
23
13
10
12
10
19
10
12
23
10
23
12
24
19
22
–
23
12
9
IIIb
IIIc
13
10
19
12
19
10
17
13
17
15
23
12
–
12
12
20
10
20
12
16
12
16
16
22
10
–
12
11
19
11
19
11
16
12
16
13
22
11
–
13
11
19
11
19
11
17
13
17
14
20
11
–
11
13
11
20
11
13
22
11
22
11
11
20
21
–
IIId
IIIe
IIIf
IV
)
11
9
)
)
20
9
)
V)
11
21
9
VIa
)
VIb
)
VIc
VId
VIe
VIf
)
21
13
23
20
21
–
)
)
)
Streptomycin
Fusidic acid
17
17
18
18
A. n.
cillus subtilis
,
Aspergillus niger
;
Pen. sp
.
,
Penicillium sp.; C. a.
,
Candida albican
; Str. sp., Streptomyces sp., R. i., Rhodotorula ingeniosa; B. s., Baꢀ
;
S. l., Streptococcus lactis
;
E. c. Escherichia coli
,
;
P. sp Pseudomonas sp.
.
,
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 40
No. 3
2014

310
AHMED A. FAYED et al.
Cl
PhNHNH
Cl
2
CH3
N
Cl
N
N
S
N
H
N
O
O
Cl
Cl
IV
OEt
X
NH2
Cl
S
Cl
N
CH3
N
O
CH3
Cl
Cl
N
N
N
S
N
O
N
X
II
IIIa–e
OEt
OEt
a, X = H;
, X = CH3;
b
, X = Cl;
c
, X = OMe;
NH NH
2
2
d
e
, X = F
Cl
Cl
Cl
S
Cl
R
NCX
N
O
CH3
NH2
N
O
CH3
Cl
N
Cl
N
R
N
N
S
NH
N
N
X
N
H
V
OEt
OEt
VIa–f
a, X = O, R = H;
, X = O; R = H;
b
e
, X = O, R = Cl;
, X = O, R = Cl;
c
, X = O, R = CH3;
d
f, X = O, R = CH3
Scheme 2. Synthetic pathway for starting material (III
–VI).
Biological Activity
The zone of inhibition (in mm) was compared with
the inhibition zones obtained using streptomycin and
fusidic as positive controls for antibacterial and antiꢀ
fungal activities, respectively. From the obtained reꢀ
sults in the table, antibacterial screening revealed that
EXPERIMENTAL
Melting points were measured using Electrotherꢀ
chi, Switꢀ
mal 9100 digital melting point apparatus (B
ü
zerland) and are uncorrected. IR spectra (
ν
, cm–1)
were recorded on a PerkinꢀElmer 1600 FTIR equipꢀ
ment (PerkinꢀElmer) in KBr discs. NMR spectra were
registered on a Jeol 5000 MHz spectrometer (Jeol, Jaꢀ
compounds (IIIf), (VIa), (VIc), (VIe) and (VIf
)
pan) in DMSOꢀ
125 MHz), and chemical shifts are reported as
d
6 (1H NMR at 500 MHz, 13C NMR at
showed the highest inhibition against all bacterial orꢀ
ganisms. Also, antifungal activity assay revealed that
the compounds (IIIe), (IV), and (VIe) exhibited highꢀ
est inhibition against fungi and yeast organisms. The
results of antibacterial activity assessment are summaꢀ
rized in the following table.
δ
, ppm,
relative to the internal standard Me4Si. The mass specꢀ
tra were registered at 70 eV with a Finnigan SSQ 7000
spectrometer (Madison, WI, USA) using EI and the valꢀ
m/z are indicated in Dalton. Elemental analyses
were performed on a PerkinꢀElmer 2400 analyzer (Perꢀ
ues of
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 40
No. 3
2014

SYNTHESIS AND ANTIMICROBIAL ACTIVITY
311
kinꢀElmer, USA) and were found within 0.4% of the 154.13, 154.72, 156.18, 157.45, 159.64, 163.75, 164.30
theoretical values. All reactions were followed by TLC
(Silica gel, Aluminum Sheets 60 F254, Merck). Starting
material (II) was prepared from the corresponding
(30 C); MS,
m/z (%): 624 (24) [M
+]. Calc. for
C30H21Cl3N4O3S (623.94): C, 57.75; H, 3.39; Cl, 17.05;
N, 8.98; S, 5.14. Found: C, 57.70; H, 3.33; Cl, 17.00;
N, 8.94; S, 5.10.
7ꢀ(2ꢀChloroꢀ6ꢀethoxypyridinꢀ4ꢀyl)ꢀ9ꢀ(2,6ꢀdichloꢀ
rophenyl)ꢀ2ꢀmethylꢀ3ꢀ(4ꢀmethylphenyl)pyrido[3',2':4,5]ꢀ
cinnamoyl derivatives (
procedures [1].
General procedure for compounds (IIIa–e) and
(IV). A mixture of 7ꢀ(2ꢀchloroꢀ6ꢀethoxypyridinꢀ4ꢀ
yl)ꢀ9ꢀ(2,6ꢀdichlorophenyl)ꢀ2ꢀmethylꢀ4 ꢀpyridoꢀ
[3',2':4,5]ꢀthieno[3,2ꢀ ][1,3]oxazinꢀ4ꢀone (II) [1]
(1 mmol) and substituted aniline derivatives, namely,
aniline, ꢀchloroꢀ, ꢀmethoxyꢀ, ꢀmethylꢀ, ꢀflouroꢀ
I) according to the reported
thienoꢀ[3,2ꢀ
mp 256–258
d
°
]pyrimidineꢀ4(3H)ꢀone (IIId). Yield 65%,
C (AcOH); IR: 1676 (C=O); 1H NMR:
H
d
1.36 (t, 3H, CH3), 2.28, 2.35 (2s, 6H, 2CH3), 3.82
(q, 2H, CH2), 6.98–7.68 (m, 9H, 7 PhꢀH + 2 pyrꢀH),
8.35 (s, 1H, pyrꢀ5'ꢀH);13C NMR: 14.08, 24.18, 25.14,
64.16, 99.95, 101.20, 116.25, 120.31, 121.36, 126.45,
128.82, 129.32, 129.65, 132.72, 133.36, 133.98, 134.42,
136.55, 145.63, 146.24, 149.82, 151.75, 154.16, 154.70,
p
p
p
p
aniline, or phenylhydrazine (1 mmol) in glacial acetic acꢀ
id (25 mL) was heated under reflux for 6 h. The reaction
mixture was concentrated under reduced pressure,
poured into ice–water, and the formed solid was filꢀ
tered, washed with water, dried, and crystallized from
the proper solvent to afford the corresponding 3ꢀsubꢀ
157.42, 159.60, 163.70, 164.33 (30 C); MS,
m/z (%):
608 (15) [
M
+]. Calc. for C30H21Cl3N4O2S (607.94):
C, 59.27; H, 3.48; Cl, 17.50; N, 9.22; S, 5.27. Found:
C, 59.21; H, 3.42; Cl, 17.44; N, 9.16; S, 5.22.
7ꢀ(2ꢀChloroꢀ6ꢀethoxypyridinꢀ4ꢀyl)ꢀ9ꢀ(2,6ꢀdichloꢀ
rophenyl)ꢀ2ꢀmethylꢀ3ꢀ(4ꢀflourophenyl)pyrido[3',2':4,5]
stituted pyrido[3',2':4,5]ꢀthieno[3,2ꢀ
rivatives (IIIa ) and (IV), respectively.
7ꢀ(2ꢀChloroꢀ6ꢀethoxypyridinꢀ4ꢀyl)ꢀ9ꢀ(2,6ꢀdichloꢀ
rophenyl)ꢀ2ꢀmethylꢀ3ꢀphenylpyrido[3',2':4,5]thienoꢀ
[3,2ꢀ ]pyrimidineꢀ4(3H)ꢀone (IIIa). Yield 82%, mp 118–
120 C (AcOH); IR: ν, 1671 (C=O); H NMR: 1.14
d]pyrimidine deꢀ
–e
thienoꢀ[3,2ꢀ
mp 216–218
d
°
]pyrimidineꢀ4(3H)ꢀone (IIIe). Yield 66%,
d
C (AcOH); IR: 1674 (C=O); 1H NMR:
1
°
1.14 (t, 3H, CH3), 2.15 (s, 3H, CH3), 3.76 (q, 2H,
CH2), 6.74–7.72 (m, 9H, 7 PhꢀH + 2 pyrꢀH), 8.44 (s,
(t, 3H, CH3), 2.32 (s, 3H, CH3), 3.76 (q, 2H, CH2),
7.06–7.72 (m, 10H, 8 PhꢀH + 2 pyrꢀH), 8.34 (s, 1H,
pyrꢀ5'ꢀH); 13C NMR: 14.15, 25.18, 64.12, 100.00,
101.21, 116.20, 120.32, 121.60, 124.32, 126.40, 128.84,
129.60, 132.75, 132.83, 133.32, 134.43, 136.54, 145.65,
146.22, 149.80, 151.75, 154.12, 154.70, 157.44, 159.66,
1H, pyrꢀ5'ꢀH); 13C NMR: 13.96, 24.89, 64.18, 99.99,
101.00, 115.62, 116.25, 120.36, 123.64, 125.32, 126.54,
128.85, 132.80, 133.34, 134.42, 136.56, 145.66, 146.25,
149.80, 151.70, 154.13, 154.73, 157.43, 158.12, 159.68,
163.80, 164.18 (29 C); MS, m/z: 612 (12) [M
+]. Calc.
163.78, 164.27 (29 C); MS, m/z (%): 594 (8) [M
+]; Calc.
for C29H19Cl3N4O2S (593.91): C, 58.65; H, 3.22;
Cl, 17.91; N, 9.43; S, 5.40. Found: C, 58.60; H, 3.18;
Cl, 17.86; N, 9.38; S, 5.34.
for C29H18Cl3FN4O2S (611.90): C, 56.92; H, 2.97;
Cl, 17.38; N, 9.16; S, 5.24. Found: C, 56.86; H, 2.92;
Cl, 17.32; N, 9.10; S, 5.20.
7ꢀ(2ꢀChloroꢀ6ꢀethoxypyridinꢀ4ꢀyl)ꢀ9ꢀ(2,6ꢀdichloꢀ
rophenyl)ꢀ2ꢀmethylꢀ3ꢀ(4ꢀnitrophenyl)pyrido[3',2':4,5]
7ꢀ(2ꢀChloroꢀ6ꢀethoxypyridinꢀ4ꢀyl)ꢀ9ꢀ(2,6ꢀdichloꢀ
rophenyl)ꢀ2ꢀmethylꢀ3ꢀ(4ꢀchlorophenyl)pyrido[3',2':4,5]
thieno[3,2ꢀ
d
]pyrimidineꢀ4(3H)ꢀone (IIIf). Yield 75%,
thieno[3,2ꢀ
d]pyrimidineꢀ4(3H)ꢀone (IIIb). Yield 66%,
mp 196–198
°
C (AcOH); IR: 1678 (C=O); 1H NMR:
mp 232–234
°
C (AcOH/H2O); IR: 1676 (C=O);
1H NMR: 1.16 (t, 3H, CH3), 2.18 (s, 3H, CH3), 3.80
(q, 2H, CH2), 7.18–7.78 (m, 9H, 7 PhꢀH + 2 pyrꢀH),
1.15 (t, 3H, CH3), 2.22 (s, 3H, CH3), 3.75 (q, 2H,
CH2), 7.24–8.12 (m, 9H, 7 PhꢀH + 2 pyrꢀH), 8.24
(s, 1H, pyrꢀ5'ꢀH); 13C NMR: 14.18, 25.13, 64.13,
100.01, 101.22, 116.23, 120.32, 121.30, 122.32, 126.46,
128.86, 132.75, 133.35, 134.45, 136.52, 138.82, 144.43,
145.65, 146.21, 149.81, 151.74, 154.13, 154.73, 157.46,
8.56 (s, 1H, pyrꢀ5'ꢀH); 13C NMR: 13.98, 24.88, 64.18,
99.95, 101.08, 116.24, 120.35, 123.56, 126.55, 128.86,
129.46, 129.76, 130.68, 132.80, 133.35, 134.45, 136.55,
145.68, 146.26, 149.81, 151.71, 154.15, 154.71, 157.45,
159.64, 163.75, 164.32 (29 C); MS,
m/z (%): 639 (12)
159.65, 163.76, 164.28 (29 C); MS,
m/z (%): 628 (18)
+]. Calc. for C29H18Cl3N5O4S (638.91): C, 54.52;
[M
+]; Calc. for C29H18Cl4N4O2S (628.36): C, 55.43;
[M
H, 2.84; Cl, 16.65; N, 10.96; S, 5.02. Found: C, 54.44;
H, 2.80; Cl, 16.60; N, 10.90; S, 4.96.
H, 2.89; Cl, 22.57; N, 8.92; S, 5.10. Found: C, 55.35;
H, 2.82; Cl, 22.51; N, 8.86; S, 5.04.
7ꢀ(2ꢀChloroꢀ6ꢀethoxypyridinꢀ4ꢀyl)ꢀ9ꢀ(2,6ꢀdichloꢀ
rophenyl)ꢀ3ꢀaminophenylꢀ2ꢀmethylpyrido[3',2':4,5]
7ꢀ(2ꢀChloroꢀ6ꢀethoxypyridinꢀ4ꢀyl)ꢀ9ꢀ(2,6ꢀdichloꢀ
rophenyl)ꢀ2ꢀmethylꢀ3ꢀ(4ꢀmethoxyphenyl)pyridoꢀ
thieno[3,2ꢀ
d
]pyrimidinꢀ4(3H)ꢀone (IV). Yield 62%,
[3',2':4,5]thieno[3,2ꢀ
d]pyrimidineꢀ4(3H)ꢀone (IIIc).
mp 178–180
°
C (DMF/H2O); IR: 3358 (NH), 1676
Yield 74%, mp 186–188
°
C (EtOH); IR: 1674 (C=O);
1
1H NMR: 1.16 (t, 3 H, CH3), 2.24 (s, 3H, CH3), 3.65
(s, 3H, OCH3), 3.76 (q, 2H, CH2), 6.68–7.65 (m, 9H,
(C=O); H NMR: 1.24 (t, 3H, CH3), 2.35 (s, 3H,
CH3), 3.78 (q, 2H, CH2), 6.68–7.71 (m, 10H,
8 PhꢀH + 2 pyrꢀH), 8.34 (s, 1H, pyrꢀ5'ꢀH), 9.12
(s, 1H, NH changeable with D2O); 13C NMR: 14.16,
7 PhꢀH + 2 pyrꢀH), 8.45 (s, 1H, pyrꢀ5'ꢀH); 13C NMR:
14.16, 25.12, 55.85, 64.16, 100.01, 101.22, 114.56,
116.23, 120.33, 122.36, 125.12, 126.41, 128.83, 132.73, 25.18, 64.14, 100.01, 101.21, 112.89, 116.20, 119.45,
133.37, 134.43, 136.52, 145.65, 146.21, 149.81, 151.73, 120.32, 126.40, 128.84, 129.60, 132.75, 133.34, 134.43,
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AHMED A. FAYED et al.
136.54, 145.64, 146.22, 149.14, 149.80, 151.73, 154.16, 7.70 (m, 9H, 7 PhꢀH + 2 pyrꢀH), 8.36 (s, 1H, pyrꢀ5'ꢀH),
154.70, 157.45, 159.66, 163.75, 164.28 (29 C); MS,
m/z
8.54, 8.62 (2s, 2H, 2NH exchangeable with D2O);
13C NMR: 14.12, 25.15, 64.16, 99.97, 101.18, 116.21,
(%): 609 (25) [
M
+]. Calc. for C29H20Cl3N5O2S (608.93):
C, 57.20; H, 3.31; Cl, 17.47; N, 11.50; S, 5.27; Found: 120.36, 123.02, 126.42, 128.92, 129.61, 130.00, 132.74,
C, 57.12; H, 3.26; Cl, 17.40; N, 11.43; S, 5.22.
3ꢀAminoꢀ7ꢀ(2ꢀchloroꢀ6ꢀethoxypyridinꢀ4ꢀyl)ꢀ9ꢀ
(2,6ꢀdichlorophenyl)ꢀ2ꢀmethylpyrido[3',2':4,5]thienoꢀ
133.36, 133.85, 134.44, 136.55, 145.65, 146.20, 149.16,
149.81, 153.46, 154.18, 154.71, 157.42, 159.65, 163.74,
164.36 (30 C); MS, (%): 652 (22) [
+]. Calc. for
C30H20Cl4N6O3S (686.40): C, 52.49; H, 2.94; Cl, 20.66;
m/z
M
[3,2ꢀ
pound (II) (1 mmol) and hydrazine hydrate (8 mmol) N, 12.24; S, 4.67. Found: C, 52.42; H, 2.90; Cl, 20.60;
in absolute ethanol (50 mL) was refluxed for 4 h. After N, 12.20; S, 4.60.
cooling, the obtained solid was collected by filtration,
dried, and crystallized from acetic acid to give the title
compound derivative ( ). Yield 78%, mp 224–226 C;
IR: 3348 (NH2), 1676 (C=O); 1H NMR: 1.25 (t, 3H, Yield 76%, mp 204–206
CH3), 2.31 (s, 3H, CH3), 3.80 (q, 2H, CH2), 4.74 (s,
2H, NH2 exchangeable with D2O), 7.18–7.72 (m, 5H, CH3), 2.10 (t, 3H, CH3), 2.28 (s, 3H, CH3), 3.77 (q,
2H, CH2), 7.02–7.68 (m, 9H, 7 PhꢀH + 2 pyrꢀH),
d]pyrimidinꢀ4(3H)ꢀone (V). A mixture of comꢀ
1ꢀ{(2ꢀChloroꢀ6ꢀethoxypyridinꢀ4ꢀyl)ꢀ9ꢀ(2,4ꢀdichloꢀ
rophenyl)ꢀ2ꢀmethylꢀ4ꢀoxopyrido[2',3':4,5]thieno[3,2ꢀ
pyrimidinꢀ3(4 )ꢀyl}ꢀ3ꢀ(4ꢀmethylphenyl)urea (VIc).
C (AcOH/H2O); IR: 3389–
d]
V
°
H
°
1
3276 (NH), 1675 (C=O); H NMR: 1.15 (s, 3H,
3 PhꢀH + 2 pyrꢀH), 8.42 (s, 1H, pyrꢀ5'ꢀH); 13C NMR:
14.08, 25.14, 64.16, 99.95, 101.20, 116.25, 120.31, 8.36 (s, 1H, pyrꢀ5'ꢀH), 8.49, 8.62 (2 s, 2H, 2 NH exꢀ
changeable with D2O); 13C NMR: 13.99, 24.12, 25.16,
64.10, 100.01, 100.98, 116.22, 120.35, 121.42, 126.41,
128.90, 129.61, 132.71, 132.86, 133.36, 134.08, 134.41,
136.53, 145.65, 146.24, 149.15, 149.80, 153.45, 154.16,
126.45, 129.65, 132.72, 133.36, 133.98, 136.55, 145.63,
146.24, 149.82, 154.16, 154.70, 157.42, 159.60, 163.70,
163.86, 164.33 (23 C); MS, (%): 534 (8) [
+ + 1].
Calc. for C23H16Cl3N5O2S (532.83): C, 51.85; H, 3.03;
m/z
M
Cl, 19.96; N, 13.14; S, 6.02. Found: C, 51.80; H, 3.00; 154.70, 157.45, 159.66, 163.75, 164.36 (31 C); MS,
Cl, 19.90; N, 13.10; S, 5.75.
General procedure for semicarbazides (VIa–c) and C, 55.91; H, 3.48; Cl, 15.97; N, 12.62; S, 4.81. Found:
m/z
(%): 666 (4) [M
+]. Calc. for C31H23Cl3N6O3S (665.98):
C, 55.86; H, 3.40; Cl, 15.90; N, 12.57; S, 4.76.
1ꢀ{(2ꢀChloroꢀ6ꢀethoxypyridinꢀ4ꢀyl)ꢀ9ꢀ(2,4ꢀdichloꢀ
rophenyl)ꢀ2ꢀmethylꢀ4ꢀoxopyrido[2',3':4,5]thieno[3,2ꢀ
thiosemicarbazides (VId–f). To a mixture of comꢀ
pound ( ) (1 mmol) and substituted isothiocyanate or
phenylisothiocyanate (1 mmol) in dry dioxane (25 mL),
triethylamine (2 mL) was added with stirring. The reacꢀ
tion mixture was heated under reflux for 10 h. The solꢀ
vent was evaporated under reduced pressure; the obꢀ
V
d]pyrimidinꢀ3(4
H
)ꢀyl}ꢀ3ꢀphenylthiourea (VId). Yield
C (AcOH); IR: 3365–3252 (NH),
68%, mp 165–167
°
1
1670 (C=O), 1228 (C=S); H NMR: 1.18 (s, 3H,
CH3), 2.15 (t, 3H, CH3), 3.76 (q, 2H, CH2), 4.28, 4.36
(2s, 2H, 2NH exchangeable with D2O), 6.86–7.65
(m, 10H, 8 PhꢀH + 2 pyrꢀH), 8.32 (s, 1H, pyrꢀ5'ꢀH);
13C NMR: 14.15, 25.18, 64.14, 100.01, 101.21, 116.20,
120.32, 124.65, 125.94, 126.40, 128.90, 129.60, 132.75,
133.34, 134.43, 136.54, 136.88, 145.64, 146.22, 149.14,
149.80, 154.16, 154.70, 157.45, 159.66, 163.75, 164.32,
tained residue was solidified with
formed was collected by filtration, washed with diethyl
ether, dried, and crystallized from the proper solvents
to afford the corresponding urea (VIa ) and thiourea
VId ) derivatives, respectively.
1ꢀ{(2ꢀChloroꢀ6ꢀethoxypyridinꢀ4ꢀyl)ꢀ9ꢀ(2,4ꢀdichloꢀ
rophenyl)ꢀ2ꢀmethylꢀ4ꢀoxopyrido[2',3':4,5]thienoꢀ
[3,2ꢀ ]pyrimidinꢀ3(4 )ꢀyl}ꢀ3ꢀphenylurea VIa). Yield
75%, mp 232–234 C (DMF/H2O); IR: 3405–3258
nꢀhexane. The solid
–c
(
–f
d
H
(
180.43 (30 C); MS, m/z (%): 668 (16) [M
+]. Calc. for
C30H21Cl3N6O2S2 (668.02): C, 53.94; H, 3.17; Cl, 15.92;
N, 12.58; S, 9.60. Found: C, 53.88; H, 3.12; Cl, 15.87;
N, 12.53; S, 9.55.
1ꢀ{(2ꢀChloroꢀ6ꢀethoxypyridinꢀ4ꢀyl)ꢀ9ꢀ(2,4ꢀdichloꢀ
rophenyl)ꢀ2ꢀmethylꢀ4ꢀoxopyrido[2',3':4,5]thieno[3,2ꢀ
°
(NH), 1671 (C=O); 1H NMR: 1.14 (s, 3H, CH3), 2.12
(t, 3H, CH3), 3.76 (q, 2H, CH2), 6.98–7.70 (m, 10H,
8 PhꢀH + 2 pyrꢀH), 8.42 (s, 1H, pyrꢀ5'ꢀH), 8.50, 8.65
13
(2s, 2H, 2NH exchangeable with D2O); C NMR:
14.15, 25.18, 64.14, 100.01, 101.21, 116.20, 120.32,
121.42, 124.08, 126.40, 128.90, 129.60, 132.75, 133.34,
134.43, 135.80, 136.54, 145.64, 146.22, 149.14, 149.80,
153.45, 154.16, 154.70, 157.45, 159.66, 163.75, 164.32
d
]pyrimidinꢀ3(4
H
)ꢀyl}ꢀ3ꢀ(4ꢀchlorophenyl)thiourea
C (EtOH); IR: 3342–
(VIe). Yield 72%, mp 256–258
°
3254 (NH), 1676 (C=O), 1234 (C=S); 1H NMR: 1.15
(s, 3H, CH3), 2.22 (t, 3H, CH3), 3.75 (q, 2H, CH2),
4.32, 4.44 (2s, 2H, 2NH exchangeable with D2O),
6.64–7.56 (m, 9H, 7 PhꢀH + 2 pyrꢀH), 8.34 (s, 1H,
pyrꢀ5'ꢀH); 13C NMR: 13.98, 25.14, 64.18, 99.99,
101.16, 116.22, 120.35, 126.45, 127.52, 128.95, 129.59,
(30 C); MS,
m/z (%): 652 (22) [M
+]. Calc. for
C30H21Cl3N6O3S (651.95): C, 55.27; H, 3.25; Cl, 16.31;
N, 12.89; S, 4.92. Found: C, 55.20; H, 3.20; Cl, 16.24;
N, 12.82; S, 4.85.
1ꢀ{(2ꢀChloroꢀ6ꢀethoxypyridinꢀ4ꢀyl)ꢀ9ꢀ(2,4ꢀdichloꢀ 130.02, 132.72, 133.35, 133.84, 135.40, 136.54, 145.64,
rophenyl)ꢀ2ꢀmethylꢀ4ꢀoxopyrido[2',3':4,5]thieno[3,2ꢀ 146.22, 149.18, 149.82, 154.20, 154.70, 157.40, 159.63,
]pyrimidinꢀ3(4 )ꢀyl}ꢀ3ꢀ(4ꢀchlorophenyl)urea (VIb). 163.75, 164.38, 180.65 (30 C); MS, (%): 703 (32)
Yield 82%, mp 158–160 C (Dioxan); IR: 3398–
d
H
m/z
M
+ + 1]. Calc. for C30H20Cl4N6O2S2 (702.46): C, 51.29;
°
[
1
3265 (NH), 1674 (C=O); H NMR: 1.17 (s, 3H,
H, 2.87; Cl, 20.19; N, 11.96; S, 9.13. Found: C, 51.22;
CH3), 2.18 (t, 3H, CH3), 3.78 (q, 2H, CH2), 6.98–
H, 2.82; Cl, 14.19; N, 11.90; S, 9.07.
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 40
No. 3
2014

SYNTHESIS AND ANTIMICROBIAL ACTIVITY
313
1ꢀ{(2ꢀChloroꢀ6ꢀethoxypyridinꢀ4ꢀyl)ꢀ9ꢀ(2,4ꢀdichloꢀ
rophenyl)ꢀ2ꢀmethylꢀ4ꢀoxopyrido[2',3':4,5]thieno[3,2ꢀ
]pyrimidinꢀ3(4 )ꢀyl}ꢀ3ꢀ(4ꢀmethylphenyl)thiourea (VIf).
Yield 65%, mp 223–225 C (AcOH); IR: 3356–3243
ACKNOWLEDGMENTS
The authors extend their appreciation to the Deanꢀ
ship of Scientific Research at King Saud University for
funding the work through the research group project
no. RGP–VPP–099.
d
H
°
(NH), 1672 (C=O), 1233 (C=S); 1H NMR: 1.18 (s,
3H, CH3), 2.12 (t, 3H, CH3), 2.30 (s, 3H, CH3), 3.76
(q, 2H, CH2), 4.24, 4.52 (2s, 2H, 2NH exchangeable
with D2O), 6.84–7.68 (m, 9H, 7 PhꢀH + 2 pyrꢀH), 8.35
REFERENCES
(s, 1H, pyrꢀ5'ꢀH); 13C NMR: 13.95, 24.14, 25.18,
64.12, 100.08, 101.08, 116.24, 120.36, 126.24, 126.40,
128.90, 129.63, 132.71, 133.36, 133.86, 134.22, 134.41,
136.53, 145.65, 146.24, 149.15, 153.45, 154.16, 154.70,
1. Hossan, A.S.M., AbuꢀMelha, H.M.A., AlꢀOmar, M.A.,
and Amr, A.E., Molecules, 2012, vol. 17, pp. 13642–
13655.
2. AlꢀMohizea, A.M., AlꢀOmar, M.A., Abdalla, M.M.,
and Amr, A.E., Int. J. Biol. Macromol., 2012, vol. 50,
pp. 171–179.
3. Khalifa, N.M., AlꢀOmar, M.A., Amr, A.E., and
Haiba, M.E., Int. J. Biol. Macromol., 2013, vol. 54,
pp. 51–56.
157.45, 159.66, 163.75, 164.36, 180.86 (31 C); MS,
m/z
(%): 682 (6) [
M
+]. Calc. for C31H23Cl3N6O2S2 (682.04):
C, 54.59; H, 3.40; Cl, 15.59; N, 12.32; S, 9.40. Found:
C, 54.52; H, 3.35; Cl, 15.52; N, 12.26; S, 9.34.
4. AlꢀHarbi, N.O., Bahashwan, S.A., Fayed, A.A.,
Aboonq, M.S., and Amr, A.E., Int. J. Biol. Macromol.
2013, vol. 57, pp. 165–173.
,
Biological Assay
5. Bahashwan, S.A., AlꢀHarbi, N.O., Fayed, A.A., Amr, A.E.,
Shadid, K.A., Alalawi, A.M., and Bassati, I.M.S., Int.
J. Biol. Macromol., 2012, vol. 51, pp. 7–17.
6. Ouf, N.H. and Amr, A.E., Monatsh. Chem., 2008,
vol. 139, pp. 579–585.
7. Amr, A.E., Mohamed, A.M., Mohamed, S.F., Abdelꢀ
Hafez, N.A., and Hammam, A.G., Bioorg. Med.
Chem., 2006, vol. 14, pp. 5481–5488.
8. AbouꢀGhalia M.H., and Amr, A.E., Amino Acids, 2004,
vol. 26, pp. 283–289.
9. Amr, A.E., Hegab, M.I., Ibrahim, A.A., and
Abdulla, M.M., Monatsh. Chem., 2003, vol. 134,
pp. 1395–1409.
The antimicrobial activities of some of the syntheꢀ
sized compounds were determined by the agar diffuꢀ
sion method as recommended by the National Comꢀ
mittee for Clinical Laboratory Standards (NCCLS)
[21]. The compounds were evaluated for antimicrobial
activity against the bacteria Streptomyces sp., Bacillus
subtilis
Pseudomonas sp. and antifungal activity against fungi
Aspergillus niger and Penicillium sp.) and yeast (Candida
, Streptococcus lactis, Escherichia coli, and
(
albican and Rhodotorula ingeniosa).
The concentrations of the tested compounds
(10
Bauer’s disk diffusion method. The sterile discs were
impregnated with 10 g/disc of the tested compound.
μg/mL) were used according to a modified Kirbyꢀ
10. Chakrabarti, J.K., Horsman, L., Hotten, T.M., Pullar, I.A.,
Tupper, D.E., and Wright, F.C., J. Med. Chem., 1980,
vol. 23, pp. 878–884.
11. Ram, V.J., Pandey, H.K., and Vlietinck, A.J., J. Heteroꢀ
cycl. Chem., 1981, vol. 18, pp. 1277–1285.
μ
Each tested compound was assayed in triplicate.
DMSO was used as a negative control and streptomyꢀ
cin and fusidic acid were used as standards. Calculated
average diameters (for triplicates) of the zone of inhiꢀ
bition (in mm) for tested samples were compared with
that produced by the standard drugs.
12. Fahmy, H.H. and ElꢀEraqy, W., Arch. Pharm. Res.
2001, vol. 24, pp. 171–178.
,
13. DeClercq, E., Anticancer Res., 1986, vol. 6, pp. 549–
556.
14. DeClercq, E., J. Med. Chem., 1986, vol. 29, pp. 156–
162.
15. Hammam, A.G., Sharaf, M., and AbdelꢀHafez, N.A.,
Ind. J. Chem., 2001, vol. 40B, p. 213.
16. Hammam, A.G., Fahmy, A.F.M., Amr, A.E., and
Mohamed, A.M., Ind. J. Chem. Sec. B., 2003, vol. 42B,
pp. 1985–1993.
17. Kumar, K.S., Chamakuri, S., Vishweshwar, P., Iqbal, J.,
and Pal, M., Tetrahedron Letters, 2010, vol. 51,
pp. 3269–3273.
18. Ola H.R.,Omaima G.S., and Ibrahim M.E., Europ. J.
Med. Chem., 2012, vol. 55, pp. 85–93.
Antimicrobial screening media included (1) PDA
medium (4 g dextrose/L potatoes extract), for fungi
cultivation; (2) Czapek Dox medium (10 g glucose,
2 g KNO3, 1 g K2HPO4, 0.5 g KCl, 0.5 g MgSO4,
and 0.05 g ferrous sulphate/L distilled water), for bacteꢀ
ria cultivation; and (3) medium 3 (10 g glucose, 5 g pepꢀ
tone, 3 g yeast extract, and 3 g malt extract), for yeast
cultivation.
CONCLUSIONS
A series of newly compounds (III)–(VI) were preꢀ
pared using citrazinic acid (2,6ꢀdihydroxyisoꢀnicoꢀ
tinic acid) as a starting material. The obtained derivaꢀ
tives were screened as antimicrobial and antifungal
agents. Some of the synthesized compounds exhibited
antibacterial and antifungal bioactivity compared with
streptomycin and fusidic acid used as reference drugs.
The other tested compounds were found to exhibit
moderate to low antibacterial activity.
19.
Pédeboscq, S., Gravier, D., Casadebaig, F., Hou, G.,
Gissot, A., De Giorgi, F., Ichas, F., Cambar, J., and
Pometan, J., Europ. J. Med. Chem., 2010, vol. 45,
pp. 2473–2479.
20. Ashour, H.M., Shaaban, O.G., Rizk, O.H., and Elꢀ
Ashmawy, I.M., Europ. J. Med. Chem., 2013, vol. 62,
pp. 341–351.
21. Furtada, G.L. and Medeiros, A.A., J. Clin. Microbiol.
1980, vol. 12, 550–556.
,
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 40
No. 3
2014