A convenient synthesis and biological activity of novel thieno[2,3-c]pyrazole compounds as antimicrobial and anti-inflammatory agents

Article abstract of DOI:10.1134/S1068162015010057

A new method for synthesizing 4-amino-3-methyl-1-phenyl-1H-5-substituted thieno[2,3-c]pyrazole was reported. The substituted groups at position 5 include carbonitrile, carboxamide, N-phenyl carboxamide, and benzoyl groups. The newly synthesized compounds

Full text of DOI:10.1134/S1068162015010057

ISSN 1068ꢀ1620, Russian Journal of Bioorganic Chemistry, 2015, Vol. 41, No. 1, pp. 97–104. © Pleiades Publishing, Ltd., 2015.
A Convenient Synthesis and Biological Activity
of Novel Thieno[2,3ꢀc]Pyrazole Compounds
as Antimicrobial and AntiꢀInflammatory Agents1
Adel M. Kamal ElꢀDean, Remon M. Zaki2, and Abdullah Y. Abdulrazzaq
Chemistry Department, Faculty of Science, Assiut University, Assiut, 71516 Egypt
Received April 3, 2014; in final form, June 3, 2014
Abstract—A new method for synthesizing 4ꢀaminoꢀ3ꢀmethylꢀ1ꢀphenylꢀ1
H
ꢀ5ꢀsubstituted thieno[2,3ꢀ
ꢀphenyl
c
]pyrazole was reported. The substituted groups at position 5 include carbonitrile, carboxamide,
N
carboxamide, and benzoyl groups. The newly synthesized compounds and their derivatives were characterꢀ
ized by elemental analysis and spectroscopy (IR, 1H NMR, and mass spectra). Furthermore, some of these
synthesized compounds were screened against various pathogenic bacterial and fungal strains. The results
demonstrate that most of the synthesized compounds possess a significant antibacterial activity against gramꢀ
positive and gramꢀnegative bacteria. In addition, most of these compounds showed a remarkable antiꢀfungal
activity. On the other hand, some of the synthesized compounds possess high antiꢀinflammatory activity,
which was demonstrated using the carrageenanꢀinduced rat paw edema assay.
Keywords: synthesis, reactions, thienopyrazole, thienopyrazolopyrimidine, thienopyrazoloimidazolopyrimidine
DOI: 10.1134/S1068162015010057
2 1
INTRODUCTION
achieve this using thiourea [36, 50–52] as it has been
reported in other molecules but all attempts failed.
Pyrazole derivatives display a broad spectrum of
After several attempts, we displaced chlorine with
sulfur in the presence of sodium borohydride and used
the product in situ, without isolation, for the next
reaction.
biological activities, such as antiꢀinflammatory [1–5],
antimicrobial [5–8], antioxidant [6], anticancer [9–11],
fungicidal [12], and antiviral activities [9–11, 13, 14].
Some pyrazole derivatives were reported to possess
high affinity and selectivity towards A2b adenosine
receptor antagonists [15]. Particularly, aryl pyrazoles
are important in medicinal and pesticidal chemistry
RESULTS AND DISCUSSION
[16]. Thieno[2,3ꢀ
of potent kinase inhibitors [17].
c]pyrazoles A are an important class
5ꢀMethylꢀ2ꢀphenylꢀ2,4ꢀdihydropyrazoleꢀ3ꢀone (
treated with Vilsmeier’s reagent afforded 5ꢀchloroꢀ3ꢀ
methylꢀ1ꢀphenylꢀ ꢀpyrazolꢀ4ꢀcarbaldehyde II).
When aldehyde (II) was allowed to react with hydroxyl
amine hydrochloride in ethanol in the presence of
sodium acetate, the corresponding 5ꢀchloroꢀ3ꢀ
I)
1H
(
Literature survey on thienopyrazole revealed that
most of papers focus on synthesis of the theinopyraꢀ
zole substituted at position 5 similar to structure
Few reports deal with ꢀbifunctionalized thienopyraꢀ
zole similar to structure [18–33].
B.
o
methylꢀ1ꢀphenylꢀ
III) was obtained. The pyrazole aldehyde oxime (III
was dehydrated using acetic anhydride into the correꢀ
1Hꢀpyrazoleꢀ4ꢀcarbaldehyde oxime
A
(
)
In continuation of our previous work in the syntheꢀ
sis of heterocyclic compounds containing pyrazole
moiety [34–49] we report a novel facile method of
sponding 5ꢀchloroꢀ3ꢀmethylꢀ1ꢀphenylꢀ1Hꢀpyrazoleꢀ
4ꢀcarbonitrile derivative (IV).
synthesis of thieno[2,3ꢀ
c]pyrazole substituted at posiꢀ
tion 4 and 5 similar to structure
A.
R2
R2
N
The difficulty of synthesis of such structures is
caused by the fact that most syntheses are based on a
starting molecule in which mercapto group is adjacent to
cyano group. In the case of pyrazole, the chlorine atom is
very difficult to displace with sulfur atom. We tried to
R3
R4
N
R3
S
N
S
N
R1
R1
A
B
1
The article is published in the original.
2
Fig. 1. The substituent positions in thieno[2,3ꢀc]pyrazole.
Corresponding author: eꢀmail: remonch2003@yahoo.com.
97

98
ADEL M. KAMAL ELꢀDEAN et al.
15
14
12
14
12
14
OH
12
H
O
12
Me
Me
3
4
6
13 N
Me
3
4
C N
3
4
Me
3
4
13
13
2
H
5
5
5
2
1
i
ii
1
iii
N
1
N
2
N
5
1
N
Cl
2
N
Cl 16
N
N
O 13
15
6
N
Cl15
6
6
11
10
7
11
7
8
7
8
11
10
7
8
11
8
10
10
9
9
9
9
(I
)
(II
)
(III
)
(IV)
v
iv
16
15
12
14
Me
Me
NH2
6
5
8
CN
Me
3
4
6
N
13
v
ii
vi
5
16
3
1
2
4 N
Me
N
1
N
CN
SH
N
2
7
N
SNa
N
S
Y
S
Y
9
15
N
14
13
10
11
11
10
7
N
8
12
9
(
VIIa–f
)
(
VIa–f
)
(C)
(V)
(
(
(
(
(
(
VI), (VIIa): Y = CN
VI), (VIIb): Y = CONH2
VI), (VIIc): Y = CONHPh
VI), (VIId): Y = CONHPhMeꢀ
VI), (VIIe): Y = CONHPhOMeꢀ
VI), (VIIf): Y = CONHPhClꢀ
p
p
p
Regents:
i
= POCl3/DMF; ii = H2NOH.HCl/AcONa/EtOH; iii = Ac2O;
i
iv = H2NCSNH2/EtOH;
= S8/NaBH4/EtOH; v = YCH2X/EtOH/AcONa; vii = EtOH/EtONa
v
Scheme 1. Synthesis of 4ꢀaminoꢀ3ꢀmethylꢀ1ꢀphenylꢀ5ꢀsubstituted 1Hꢀthieno[2,3ꢀc]pyrazoles (VIIa–f).
The attempt to synthesize thieno[2,3ꢀ
through converting chloropyrazole carbonitrile (IV
into mercaptopyrazole ( ) using thiourea in ethanol,
as with other moieties, followed by reacting with
ꢀhalogenated compounds, failed. This forced us to
c
]pyrazole
(VI), and appearance of the signal at 6.90, characterisꢀ
)
tic for NH2. Also, the IR of (VIIb) showed disappearance
of the band characteristic for CN group at 2220 cm–1 in
compound (VIb) and appearance of bands characterꢀ
istic for NH2 group. Mass spectrum of compound
V
α
search for another method to synthesize thienopyraꢀ
zole (VII). The desired result was achieved by the reacꢀ
tion of elemental sulfur with chloropyrazole in the
presence of sodium borohydride through reduction of
sulfur in ethanol to afford not isolated intermediate
(
VIb) showed molecular ion peak at 272.14 and
showed the base peak at 255.77, which means that the
molecular ion lost one molecule of ammonia to give
the base peak.
sodium salt
tion with
lated mercaptopyrazole carbonitrile (VIa
pounds (VIa ) underwent ThorpeꢀZiegler cyclizaꢀ
tion upon heating in ethanolic sodium ethoxide
solution to afford thienopyrazole (VIIa ). Converꢀ
C
, which was used in situ in the next reacꢀ
ꢀhalogenated compound to afford ꢀalkyꢀ
). Comꢀ
When aminocarboxamide compound (VIIb) was
allowed to react with triethyl orthoformate in presence
of catalytic amount of acetic acid, 3ꢀmethylꢀ1ꢀpyraꢀ
zolothienopyrimidinone (VIII) was obtained.
α
S
–f
–
f
On the other hand, the reaction of (VIIb) with
chloroacetyl chloride in dioxane followed by neutralꢀ
ization with sodium carbonate solution afforded the
–f
sion of (VIb) to (VIIb) was proven by spectral analysis,
1H NMR revealed the disappearance of the signal at
3.30, characteristic for –CH2– group in compound chloroacetyl amino compound (IX).
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 41
No. 1
2015

SYNTHESIS AND BIOLOGICAL ACTIVITY OF NOVEL THIENO[2,3ꢀ
NH2
c
]PYRAZOLES
99
Cl
H
Me
Me
Me
N
N
NH2
O
i
ii
N
N
NH
N
NH2
S
N
S
N
N
S
O
Ph
O
Ph
Ph
O
(
VIII
)
(
VIIb
)
(IX)
Reagents:
i
= HC(OEt)3/AcOH, ii = ClCH2COCl/dioxane
Scheme 2. Reaction of aminothieno[2,3ꢀc]pyrazole carboxamide (VIIb) with triethylorthoformate and chloroꢀ
acetyl chloride.
Antimicrobial Activity
compound (VIIe) and 4ꢀaminoꢀ
ꢀmethylꢀ1ꢀphenylꢀ ꢀthieno[2,3ꢀ
boxamide compound (VIIf) were high compared with
other tested compounds, which did not affect any
strain. Chloroacetylation of aminocaboxamide (VIIb
afforded chloroacetylamino derivative (IX) showing
high antibacterial activity against gramꢀpositive bacteꢀ
ria, such as Staphylococcus aureus and Clostridium difꢀ
ficile. Also, compound (IX) showed moderate activity
against Streptococcus strains, which are gramꢀpositive
bacteria, whereas it showed low antibacterial activities
against all tested strains of gramꢀnegative bacteria.
N
ꢀ(4ꢀchlorophenyl)ꢀ
3
1H
c]pyrazoleꢀ5ꢀcarꢀ
Some of the synthesized compounds were chosen
and screened in vitro for their antimicrobial activity
against some strains of bacteria and fungi. Antifungal
and antibacterial activities of tested compounds were
evaluated by the reported method using 0.005%
)
(
50 µg/mL) concentration of selected compounds in
DMSO as a solvent. The inhibition zone (mm) for the
antifungal activity was compared with clotrimazole as
a reference. In the case of antibacterial activity, conꢀ
centration of tested compounds was 2% and the inhiꢀ
bition zone in mm was compared with a series of antibiꢀ
otics according to the sensitivity of each type of bacteria
to the most effective antibiotic for it as a reference.
Some tested compounds showed high antibacterial
On the other hand, compounds (VIIe) and (VIIf
showed high antifungal activity against Candida albiꢀ
cans Trichophyton rubrum, and Aspergillus flavus
)
,
,
activity (Table 1). Aminocarbonitrile compound (VIIa
)
while compound (VIIb) showed the highest antifungal
activity against Trichophyton rubrum (Table 2). Howꢀ
ever, compounds (VIIa), (VIIb), and (VIIe) showed
moderate activity against some species of fungi, while
compound (IX) did not affect any of fungi species
tested. Compound (VIIa) was inactive with respect to
most of the tested species of fungi.
show low antibacterial activity compared with other
tested compounds. We found that aminocarboxamide
compound (VIIb) showed high antibacterial activity
against some strains of gramꢀnegative bacteria, such as
Esherichia coli and Klebsiella pneumoniae, with values
almost the same as those of the corresponding referꢀ
ence antibiotics (nitrofuratoin and imipenem, respecꢀ
tively). Also, compound (VIIb) showed moderate
activity against most of the tested strains of gramꢀposꢀ
AntiꢀInflammatory Activity
itive bacteria and against Pseudomonas aeruginosa
which is a gramꢀnegative bacterium. The antibacterial
activities of 4ꢀaminoꢀ ꢀ(4ꢀchlorophenyl)ꢀ3ꢀmethylꢀ
1ꢀphenylꢀ ꢀthieno[2,3ꢀ ]pyrazoleꢀ5ꢀcarboxamide
,
The results of antiꢀinflammatory activity assessꢀ
ment for some of the synthesized compounds are preꢀ
sented in Figs. 2 and 3.
N
1
H
c
Table 1. Antibacterial activity (inhibition zone, mm)
Reference
antibacterial
agent
Ref
Compound
(
VIIa
)
(
VIIb
)
(
VIIe
)
(
VIIf
)
(IX)
Bacteria strain
value sensitivity
Staphylococcus aureus (+)
Streptococcus pneumoniae (+)
Clostridium difficile (+)
Esherichia coli (–)
Klebsiella pneumoniae (–)
Pseudomonas aeruginosa (–)
R (8)
R (7)
R (8)
R (7)
R (7)
R (8)
I (14)
I (13)
I (14)
S (15)
S(16)
S(17)
S(17) S (16) Levofoxacin
S (16) I (13) Ciprofloxacin
S (15) S (21) Ofloxacin
S
S
S
S
S
S
23
28
22
21
23
20
S (21) S (20) S (20) I (10)
S (22) S (22) S (18) I (9)
I (14)
Nitrofuratoin
Imipenem
Clindamycin
S (19) S (20) I (9)
The amount added in each pore, 50 mL. S means sensitive, I means intermediate sensitivity, and R means resistant. (+) indicates gramꢀ
positive bacteria and (–), gramꢀnegative ones.
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 41
No. 1
2015

100
ADEL M. KAMAL ELꢀDEAN et al.
*
0.75
0.50
0.25
0
*
*** **
**
***
***
Ref.
*
*
(VIIa)
(VIIb)
*
(VIIe)
(VIIf)
(VIII)
30
60
120 180
Time, min
240
300
Fig. 2. Paw edema inhibition by compounds (VIIa), (VIIb), (VIIe), (VIIf), (VIII), and the reference compound, indomethacin.
The inflammatory response is represented as the cant difference from indomethacin, while the effect of
amino carboxamide compound (VIIb) was signifiꢀ
cantly different ( < 0.01). After 5 h, aminomethoxꢀ
yphenyl carboxamide (VIIe) and aminochlorophenyl
carboxamide (VIIf) persisted in showing no significant
difference from indomethacin, which means they are
potent antiꢀinflammatory agents.
percentage of the increase in paw swelling upon the
antiꢀinflammatory effect of tested compounds (Fig. 2)
and as the percentage of paw edema inhibition (Fig. 3)
calculated for each group at each time point using the
following ratio:
P
(Vt – V0)control – (Vt – V0)treated × 100/(Vt –V0)control,
In this work we synthesized bifunctional
thieno[2,3ꢀc]pyrazole compounds by incorporating
where Vt is the average volume of paw swelling at a
given time point and V0 is the average volume just after
carrageenan injection.
sulfur metal instead of chlorine in 5ꢀchloroꢀ3ꢀmethylꢀ
1ꢀphenylꢀ ꢀpyrazoleꢀ4ꢀcarbonitrile compound (IV)
1H
by using sulfur metal in the presence of sodium boroꢀ
hydride followed by Thorpe–Ziegler cyclization. The
novel synthesized compounds showed high antiꢀ
microbial and antiꢀinflammatory activities.
From the previous results we found that some of the
tested compounds showed high antiꢀinflammatory
activity compared with indomethacin. For example, a
significant difference exists between indomethacin
and aminocarboxamide (VIIb) with the
P value less
EXPERIMENTAL
than 0.05, which means that compound (VIIb) did not
affect the inflammation in rats after 1 h of treatment.
After 2 h, most compounds showed significant differꢀ
All melting points are corrected and measured on a
FisherꢀJohn apparatus. IR spectra (KBr,
recorded with a PerkinꢀElmer 1430 Spectrophotometer.
ν
, cm–1) were
ences from indomethacin. Effect of compound (VIIf
was the closest to that of indomethacin. After 3 h, amiꢀ
nomethoxyphenyl carboxamide (VIIe) and amiꢀ
)
13
1H NMR and C NMR spectra (
δ, ppm; J, Hz) were
obtained on a Varian EMꢀ390 MHz (90 MHz) and
nochlorophenyl carboxamide (VIIf) show no signifiꢀ Joel (400 MHz) spectrometers in CDCl3, DMSOꢀd6
,
Table 2. Antifungal activity (inhibition zone, mm)
Compound
Fungal strains
Ref
(
VIIa
)
(
VIIb
)
(
VIIe
)
(
VIIf
)
(IX)
value
sensitivity
Candida albicans
Trichophyton rubrum
Aspergillus flavus
Fusarium oxysporum
Scopulariopsis brevicaulis
Geotrichum candidum
I (10)
I (13)
R (0)
I (10)
R (0)
R (0)
I (14)
S (25)
R (0)
R (7)
R (0)
R (8)
I (14)
S (15)
I (13)
R (8)
R (0)
R (9)
S (16)
S (18)
I (12)
R (8)
R (7)
R (0)
R (9)
R (9)
R (8)
R (8)
R (0)
R (0)
S
S
S
S
S
S
23
28
22
21
23
20
The amount added in each pore, 50 mL. Ref = clotrimazole S means sensitive, I means intermediate sensitivity, and R means resistant.
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 41 No. 1 2015

SYNTHESIS AND BIOLOGICAL ACTIVITY OF NOVEL THIENO[2,3ꢀ
c]PYRAZOLES
101
%
100
(VIIa)
75
50
25
0
(VIIb)
(VIIe)
(VIIf)
(VIII)
30
60
120 180
Time, min
240
300
Fig. 3. Paw edema inhibition by compounds (VIIa), (VIIb), (VIIe), (VIIf), and (XIII), % to the reference compound (indomethacin).
and CF3CO2D using Me4Si as internal standard. Mass cipitate which formed by stirring was filtered off,
spectra were measured on a JoelꢀJMS 600 spectromeꢀ
ter. Analytical data were obtained on Elemental Analyze
system GmbHꢀVarioEL V.3 microanalyzer in the central
lab of Assiut University. Compounds (I)–(IV) were synꢀ
thesized according to literature procedure [53].
dried, and recrystallized from ethanol as white crysꢀ
tals. Yield 10 g (80%); mp 144–146°C. IR spectrum:
3450, 3300 (NH2), 3050 (CH aromatic), 2920, 2850
(CH aliphatic), 2220 (CN), 1660 (CO amide), 1590
(C=N). 1H NMR (300 MHz, DMSOꢀ
d6): 2.35 (s, 3H,
CH3), 3.30 (s, 2H, CH2), 7.15 (s, 2H, NH2 disapꢀ
peared by D2O), 7.30–7.70 (m, 5H, ArH). EIꢀMS:
m/z 272.14 [M
+]. Found: C, 57.26; H, 4.50; N, 20.60;
S, 12.00%. Calcd.: C13H12N4OS (272.33): C, 57.34;
H, 4.44; N, 20.57; S, 11.77%.
3ꢀMethylꢀ1ꢀPhenylꢀ5ꢀSubstituted Thiopyrazoleꢀ4ꢀ
Carbonitrile (VIa–f). General Procedure
Sodium borohydride (4 g, 0.105 mol) was added to
a suspension of finely powered sulfur (4 g, 0.125 mol)
in absolute ethanol (60 mL) in an ice bathin small porꢀ
tions till all sulfur powder dissolved, then chlorocyanꢀ
opyrazole (IV) (10 g, 46 mmol) was added to the reacꢀ
tion mixture with stirring in ice bath for 1 h. The reacꢀ
tion mixture was refluxed for 4 h followed by cooling.
2ꢀ(4ꢀCyanoꢀ3ꢀmethylꢀ1ꢀphenylꢀ1
ꢀphenylacetamide (VIc) was obtained by the reaction
with 2ꢀchloroꢀ ꢀphenylacetamide. The solid precipiꢀ
Hꢀpyrazolꢀ5ꢀylthio)ꢀ
N
N
tate formed was filtered off, dried, and recrystallized
from ethanol as white crystals. Yield: 11 g (69%); mp
The
αꢀhalogenated alkylating agent (46 mmol) was
128–130
matic), 2920, 2885 (CH aliphatic), 2227 (CN), 1654
(CO). 1H NMR (400 MHz, DMSOꢀ
6): 2.56 (s, 3H,
°C. IR spectrum: 3160 (NH), 3057 (CH aroꢀ
added to the mixture. The reaction mixture was left
overnight with stirring. The solid precipitate which
formed on cold was filtered off, dried, and recrystalꢀ
lized from the proper solvent.
d
CH3), 4.03 (s, 2H, CH2), 7.08–7.52 (m, 9H, ArH),
10.15(s, H ,NH). Found: C, 65.72; H, 4.55; N, 15.95;
S, 9.40%. Calcd.: C19H16N4OS (348.43): C, 65.50; H,
4.63; N, 16.08; S, 9.20%.
5ꢀ(Cyanomethylthio)ꢀ3ꢀmethylꢀ1ꢀphenylꢀ1Hꢀpyraꢀ
zoleꢀ4ꢀcarbonitrile (VIa) was obtained by the reaction
of chlorocyanopyrazole (IV) with chloroacetonitrile as
described above. The solid precipitate was recrystalꢀ
lized from ethanol as white crystals. Yield 10 g (85%);
2ꢀ(4ꢀCyanoꢀ3ꢀmethylꢀ1ꢀphenylꢀ1
ꢀ( ꢀtolyl)ꢀacetamide (VId) was obtained by the reacꢀ
tion with 2ꢀchloroꢀ ꢀ( ꢀtolyl)ꢀacetamide. The solid
Hꢀpyrazolꢀ5ꢀylthio)ꢀ
N
p
mp 78–80
2985–2925 (CH aliphatic), 2275, 2227 (2CN).
1H NMR spectrum (90 MHz, DMSOꢀ
6): 2.85 (s,
3H, CH3), 3.85 (s, 2H, CH2), 7.60–7.40 (m, 5H,
ArH). EIꢀMS: . Found: C, 61.23; H,
254.40 [M+
°C. IR spectrum: 3035 (CH aromatic),
N
p
precipitate formed was filtered off, dried, and recrysꢀ
tallized from ethanol as white crystals. Yield: 10.5 g
d
(63%); mp 126–128°C. IR spectrum: 3130 (NH),
3070 (CH aromatic), 2905, 2885 (CH aliphatic), 2220
m/z
]
4.20; N, 21.98; S, 12.75%. Calcd.: C13H10NO4S
(254.32): C, 61.40; H, 3.96; N, 22.03; S, 12.61%.
1
(CN), 1660 (CO). H NMR (400 MHz, DMSOꢀd6):
2.31 (s, 3H, CH3ꢀPh), 2.56 (s, 3H, CH3), 4.03 (s, 2H,
CH2), 7.21–7.52 (m, 9H, ArH), 9.21(s, H, NH).
chloroacetamide as described above. The solid preꢀ Found: C, 66.05; H, 4.95; N, 15.35; S, 8.65%. Calcd.:
2ꢀ(4ꢀCyanoꢀ3ꢀmethylꢀ1ꢀphenylꢀ1Hꢀpyrazolꢀ5ꢀylthio)ꢀ
acetamide (VIb) was obtained by the reaction with
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 41
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102
ADEL M. KAMAL ELꢀDEAN et al.
C20H18N4OS (362.46): C, 66.28; H, 5.01; N,15.46; S, 2H, NH2), 7.30–7.70 (m, 5H, ArH). EIꢀMS:
8.85%.
2ꢀ(4ꢀCyanoꢀ3ꢀmethylꢀ1ꢀphenylꢀ1
ꢀ( ꢀanisyl)ꢀacetamide (VIe) was obtained by the reꢀ
action with 2ꢀchloroꢀ ꢀ( ꢀanisyl)ꢀacetamide. The
N,1ꢀdiphenylꢀ1Hꢀthieno[2,3ꢀ
m/z
271.60 [M
+ – 1]. Found: C, 57.44; H, 4.55; N, 20.47;
S, 11.65%. Calcd.: C13H12N4OS (272.33): C, 57.34;
H, 4.44; N, 20.57; S, 11.77%.
Hꢀpyrazolꢀ5ꢀylthio)ꢀ
N
p
N
p
4ꢀAminoꢀ3ꢀmethylꢀ
]pyrazoleꢀ5ꢀcarboxamide (VIIc) was obtained from
compound (VIc) as white crystals in 68% yield. mp
202–204 C; IR spectrum: 3304, 3201 (NH2), 3135
(NH), 3050 (CH aromatic), 2007, 2887 (CH aliphatꢀ
ic), 1686 (CO); 1H NMR (400 MHz, DMSOꢀ
6): 2.63
solid precipitate formed was filtered off, dried, and reꢀ
crystallized from ethanol as white crystals. Yield: 13 g
c
(74%); mp 148–150
3035 (CH aromatic), 2976, 2870 (CH aliphatic), 2229
(CN), 1655 (CO ). 1H NMR (400 MHz, DMSOꢀ
6):
°C. IR spectrum: 3304 (NH),
°
d
d
2.52 (s, 3H, CH3), 3.70 (s, 2H, CH2), 7.30–8.00 (m,
9H, ArH), 9.15 (s, 1H, NH). Found: C, 63.55; H,
4.70; N, 14.77; S, 8.32%. Calcd.: C20H18N4OS
(378.46): C, 63.47; H, 4.79; N,14.80; S, 8.47%.
(s, 3H, CH3), 6.64 (s, 2H, NH2), 7.10–8.03 (m, 9H,
ArH), 8.89 (s, H, NH). Found: C, 65.35; H, 4.55; N,
15.95; S, 9.40%. Calcd.: C19H16N4OS (348.43): C,
65.50; H, 4.63; N,16.08; S, 9.20%.
2ꢀ(4ꢀCyanoꢀ3ꢀmethylꢀ1ꢀphenylꢀ1
ꢀ( ꢀchlorophenyl)ꢀacetamide (VIf) was obtained by
the reaction with 2ꢀchloroꢀ ꢀ(4ꢀchlorophenyl)ꢀaceꢀ
tamide. The solid precipitate formed was filtered off,
dried, and recrystallized from ethanol as white crysꢀ
Hꢀpyrazolꢀ5ꢀylthio)ꢀ
4ꢀAminoꢀ3ꢀmethylꢀ1ꢀphenylꢀNꢀpꢀtolyl
[2,3ꢀ ]pyrazoleꢀ5ꢀcarboxamide (VIId) was obtained
from compound (VId) as white crystals in 70% yield.
mp 208–210 C; IR spectrum: 3290, 3215 (NH2),
3130 (NH), 3070 (CH aromatic), 2905, 2885 (CH aliꢀ
phatic), 1655 (CO); H NMR (400 MHz, DMSOꢀd6):
2.30 (s, 3H, CH3ꢀPh), 2.63 (s,3H,CH3), 6.64 (s, 2H,
NH2), 7.30–7.58 (m, 9H, ArH), 8.84 (s, H, NH).
EIꢀMS: m/z 361.20 [M
+]. Found: C, 66.07; H, 4.95;
N, 15.35; S, 8.65%. Calcd.: C20H18N4OS (362.46): C,
66.28; H, 5.01; N, 15.46; S, 8.85%.
ꢀ1Hꢀthieno
N
p
c
N
°
tals. Yield: 13 g (74%); mp 132–134°C. IR spectrum:
3304 (NH), 3035 (CH aromatic), 2976, 2870 (CH alꢀ
1
1
iphatic), 2229 (CN), 1655 (CO). H NMR (400
MHz, DMSOꢀd6): 2.32 (s, 3H, CH3), 3.70 (s, 2H,
CH2), 7.33–7.99 (m, 9H, ArH), 10.28 (s, 1H, NH).
Found: C, 59.83; H, 3.90; Cl, 9.20; N, 14.70; S,
8.50%. Calcd.: C19H15ClN4OS (382.87): C, 59.60; H,
3.95; Cl,9.26; N,14.63; S, 8.37%.
4ꢀAminoꢀ3ꢀmethylꢀ1ꢀphenylꢀNꢀpꢀanisyl
[2,3ꢀ ]pyrazoleꢀ5ꢀcarboxamide (VIIe) was obtained
from compound (VIe) as white crystals in 65% yield.
mp 208–210 C; IR spectrum: 3429, 3329, 3275
(2NH2, NH), 3030 (CH aromatic), 2860 (CH aliphatic),
1625 (CO); 1H NMR (400 MHz, DMSOꢀ
6): 2.48 (s,
ꢀ1Hꢀthieno
c
4ꢀAminoꢀ3ꢀMethylꢀ5ꢀSubstitutedꢀ1ꢀPhenylꢀ1Hꢀ
Thieno[2,3ꢀc]ꢀ5ꢀCarbonitrilepyrazole (VIIa–f).
General Procedure
°
d
A mixture of a substituted pyrazole carbonitrile
compound (VIa–f) (4 g, 0.016 mol) in absolute ethaꢀ
nol (20 mL) and sodium ethoxide solution (2.5 mL) was
gently refluxed for 10 min. The solid precipitate which
formed on hot was filtered off, dried, and recrystallized
from the mixture of ethanol–dioxane, 2 : 1.
4ꢀAminoꢀ3ꢀmethylꢀ1ꢀphenylꢀ1Hꢀthieno[2,3ꢀc]pyraꢀ
zoleꢀ5ꢀcarbonitrile (VIIa) was obtained from comꢀ
pound (VIa) as white crystals in 75% yield. mp 198–
3H, CH3), 3.71 (s, 3H, OCH3), 7.15 (s, 2H, NH2),
7.30–7.70 (m, 9H, ArH), 9.15 (s, H, NH). Found:
C, 63.55; H, 4.70; N, 14.73; S; 8.37%. Calcd.:
C20H18N4O2S (378.46): C, 63.47; H, 4.79; N, 14.80;
S, 8.47%.
4ꢀAminoꢀ3ꢀmethylꢀ1ꢀphenylꢀNꢀpꢀchlorophenyl
thieno[2,3ꢀ ]pyrazoleꢀ5ꢀcarboxamide (VIIf) was obꢀ
tained from compound (VIf) as white crystals in 70%
yield. mp 204–206 C; IR spectrum: 3429, 3336, 3275
(2NH2, NH), 3050 (CH aromatic), 2880 (CH aliphatꢀ
ic), 1640 (CO); 1H NMR (400 MHz, DMSOꢀ
6): 2.49
(s, 3H, CH3), 7.25 (s, 2H, NH2), 7.31–7.71 (m, 9H,
ArH), 9.37 (s, H, NH). EIꢀMS: 381.30 [
+ – 1].
ꢀ1Hꢀ
c
°
200
(CH aromatic), 2950, 2890 (CH aliphatic), 2184
(CN); 1H NMR (400 MHz, DMSOꢀ
6): 3.37 (s, 3H,
CH3), 6.93 (s, 2H, NH2), 7.26–7.56 (m, 5H, ArH).
EIꢀMS: 254.33 [
+]. Found: C, 61.50; H, 4.03;
°C; IR spectrum: 3456, 3359, 3200 (NH2), 3045
d
d
m/z
M
m/z
M
Found: C, 59.71; H, 4.05; Cl, 9.04; N, 14.55; S,
8.32%. Calcd.: C19H15NClN4OS (382.87): C, 59.60; H,
3.95; Cl, 9.26; N, 14.63; S, 8.37%.
N, 21.99; S, 12.65%. Calcd.: C13H10N4S (254.32): C,
61.40; H, 3.96; N, 22.03; S, 12.61%.
4ꢀAminoꢀ3ꢀmethylꢀ1ꢀphenylꢀ1Hꢀthieno[2,3ꢀc]pyraꢀ
zoleꢀ5ꢀcarboxamide(VIIb) was obtained from comꢀ
pound (VIb) as white crystals in 70% yield. mp 214–
3ꢀMethylꢀ1ꢀphenylpyrazolo[3',4':5,4]thieno[3,2ꢀ
d]
pyrimidinꢀ7(6 )ꢀone (VIII). To a refluxed mixture of
H
amino carboxamide (VIIb) (2 g, 7 mmol) and triethyl
orthoformate (6 mL, 36 mmol), few drops of glacial
acetic acid was added. The reaction mixture was heatꢀ
ed under reflux for 30 min. The solid precipitate which
is formed on hot was filtered off, dried, and recrystalꢀ
216
(CH aromatic), 2910 (CH aliphatic), 1635 (CO
amide), 1580 (C=N); 1H NMR (90 MHz, DMSOꢀ
6):
2.60 (s, 3H, CH3), 6.90 (s, 2H, NH2 amide), 7.0 (s,
°C; IR spectrum: 3400, 3305, 3190 (2NH2), 3050
d
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 41
No. 1
2015

SYNTHESIS AND BIOLOGICAL ACTIVITY OF NOVEL THIENO[2,3ꢀ
c]PYRAZOLES
103
ized from dioxane as white needles. Yield: 1.8 g (86%); of 1% freshly prepared solution of carrageenan in disꢀ
tilled water into the rightꢀhind paws of each rat of all
the groups. Animals of group A/B/C, were treated
with the single dose of tested compound, group D was
treated with indomethacin. Paw thickness were meaꢀ
sured just before the carrageenan injection, that is, at
“0 hour” and then at 30 min, 1, 2, 3, 4, and 5 h after
carrageenan injection. Increase in paw thickness was
measured as the difference in paw thickness at
“0 hour” and paw thickness at relevant time point.
mp 350–352°C; IR spectrum: 3380 (NH broad), 3050
(CH aromatic), 3000, 2900 (CH aliphatic), 1665
(CO amide), 1590 (C=N); H NMR (400 MHz,
DMSOꢀd6): 2.50 (s, 3H, CH3), 7.30–7.80 (m, 5H,
ArH), 8.30 (s, 1H, CH pyrimidine), 10.50 (s, 1H,
NH), Found: C, 59.44; H, 3.60; N, 19.60 ; S, 11.00%.
Calcd.: C14H10N4OS (282.33) C; 59.56, H; 3.57, N;
19.84 ,S; 11.36%.
1
4ꢀ(2ꢀChloroꢀacetylamino)ꢀ3ꢀmethylꢀ1ꢀphenylꢀ
1Hꢀthieno[2,3ꢀc]pyrazoleꢀ5ꢀcarboxamide (IX). A soꢀ
Statistical Analysis
lution of compound (VIIb) (2 g, 7 mmol) and chloroꢀ
acetyl chloride (0.8 mL, 0.9 mmol) in dioxane (30 mL)
was heated on water bath for 3 hours. The solid prodꢀ
uct obtained by cooling and pouring on diluted sodiꢀ
um carbonate solution was filtered off, dried, and reꢀ
crystallized from ethanol into smoke white crystals.
The results were analyzed by one way analysis of
variance (ANOVA) followed by Newman–Keuls mulꢀ
tiple comparison test as a postꢀTest. These analyses
were carried out using computer Prism program for
windows, version 3.0 (Graph pad software, Inc., San
Diego, CA, US). The significant differences between
Yield: 2.1 g (86%); mp 258–260°C; IR spectrum: 3380,
3300, 3180 (NH, NH2), 3050 (CH aromatic), 2920, 2850 groups were accepted at
p < 0.05*, 0.01**, or
0.001***, and the data are expressed as mean stanꢀ
dard error (SE).
1
(CH aliphatic), 1690, 1650 (2C=O amide); H NMR
(400 MHz, DMSOꢀd6 2.40 (s, 3H, CH3), 4.50 (s, 2H,
):
CH2), 7.30–7.80 (m, 7H, ArH + NH2), 10.50 (s, 1H,
NH). Found: C; 51.52, H; 3.90, Cl; 10.00; N, 15.95; S,
9.00%. Calcd.: C15H13ClN4O2S (348.81): C; 51.65, H;
3.76; Cl; 10.16 N; 16.06; S, 9.19%.
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