Synthesis and Anticancer Activity of Some New Fused Pyrazoles and Their Glycoside Derivatives

Article abstract of DOI:10.1002/jhet.3208

4-(4-Chlorobenzylidene)-2,5-diphenyl-2,3-dihydro-3H-pyrazol-3-one 3a and 4-(3,4-dimethoxybenzylidene)-5-phenyl-2,3-dihydro-3H-pyrazol-3-one 3b were prepared and were reacted with phenylhydrazine, thiosemicarbazide, hydroxylamine hydrochloride, ethyl aceto

Full text of DOI:10.1002/jhet.3208

Month 2018
Synthesis and Anticancer Activity of Some New Fused Pyrazoles and
Their Glycoside Derivatives
a
b
c
*
Ibrahim F. Nassar, Ahmed F. El Farargy, and Fathy M. Abdelrazek
aFaculty of Specific Education, Ain Shams University, 365 Ramsis Street, Abassia, Cairo, Egypt
bDepartment of Chemistry, Faculty of Science, Zagazig University, Zagazig, Egypt
cDepartment of Chemistry, Faculty of Science, Cairo University, Giza 12613, Egypt
*E-mail: prof.fmrazek@gmail.com
Received January 6, 2018
DOI 10.1002/jhet.3208
Published online 00 Month 2018 in Wiley Online Library (wileyonlinelibrary.com).
4-(4-Chlorobenzylidene)-2,5-diphenyl-2,3-dihydro-3H-pyrazol-3-one 3a and 4-(3,4-dimethoxybenzy-
lidene)-5-phenyl-2,3-dihydro-3H-pyrazol-3-one 3b were prepared and were reacted with phenylhydrazine,
thiosemicarbazide, hydroxylamine hydrochloride, ethyl acetoacetate, diethylmalonate, malononitrile, ethyl
cyanoacetate, and thiourea yielding fused pyrazole derivatives. Some of the new compounds were reacted
with cyclic and acyclic sugars to produce new S-, O-, and N-glycoside derivatives. The antitumor activity
against the human breast cancer cells (MCF-7) was assessed. Four of the new compounds showed IC50
values less than those of the positive control, indicating that these four compounds are better anticancer
agents than doxorubicin.
J. Heterocyclic Chem., 00, 00 (2018).
INTRODUCTION
activity and safety advantages. As a consequence, much
attention has been paid to the design and synthesis of
fused pyrazole derivatives [18–22]. Pyrazolopyrimidines
derivatives are known to have potent pharmacological
activities that enabled using them as anticancer agents
[23], GSK-3 inhibitors [24], and antiviral agents [25].
They also manifest potential cytotoxicity activity against
human laryngeal epidermoid carcinoma cells (Hep2) [26].
Furthermore, some pyrazolo[3,4-d]pyrimidine derivatives
manifested significant activity as anti-inflammatory agents
[27]. We have been aiming in this work to synthesize
some new substituted heterocyclic systems on the basis of
pyrazole moiety of expected biological interest and to test
them as biodegradable agrochemicals [28–41].
Pyrazole derivatives represent an important class of
heterocycles owing to their prominent biological and
pharmacological activities. Several pyrazole compounds
have been reported to be potential therapeutic agents
for the treatment of inflammation [1–4] including the
marketed selective COX-2 drug, celecoxib, that have been
shown to be well tolerated with reduced gastrointestinal
side effects [5]. Moreover, various substituted pyrazoles
were reported to possess antitumor activities [6,7]; others
were used for treating Alzheimer’s disease [8] and
acquired immunodeficiency syndrome [9]. Some pyrazole
derivatives were also found to have antimicrobial [10,11],
antiviral [12], and antimalarial activities [13]. Many
pyrazole derivatives are known to exhibit a wide range of
biological properties such as cannabinoid hCB1 and hCB2
receptors, anti-inflammatory agents, inhibitors of p38
kinase, and CB1 receptor antagonists [14–17]. The
incorporation of heterocyclic rings into prospective
pharmaceutical candidates is a major strategy to obtain
RESULTS AND DISCUSSION
Chemistry.
4-(4-Chlorobenzylidene)-2,5-diphenyl-2,
3-dihydro-3H-pyrazol-3-one 3a [42] and 4-(3,4-
dimethoxybenzylidene)-5-phenyl-2,3-dihydro-3H-pyrazol-
© 2018 Wiley Periodicals, Inc.

I. F. Nassar, A. F. El Farargy, and F. M. Abdelrazek
Vol 000
3-one 3b were synthesized by Claisen–Schmidt
condensation reaction of the pyrazolonone derivatives 1a
with 4-chlorobenzaldehyde 2a or 1b with 3,4-
dimethoxybenzaldehyde 2b in sodium ethoxide solution,
respectively. The IR spectrum of 3b showed strong
absorption bands assigning the aromatic (C–H) and
pyrazole derivative 4. The structure of 4 was elucidated
by different spectral and elemental analyses (cf. section).
The IR spectrum of 4 revealed a characteristic band at
1
3150 cmꢀ1 for NH groups. On the other hand, H NMR
spectrum of 4 showed three singlet signals at 5.1, 8.5,
and 9.50 ppm for CH and two exchangeable NH groups
of pyrazole ring. The 13C NMR spectrum of 4 showed
signals at δ 56.1 and 56.2 for two methoxy groups, 76
(C4) and 101 (C-8), and at 112–154 ppm assignable for
(Ar–C, C-3, and C-7).
carbonyl group at νmax 3045 and 1665 cmꢀ1
,
respectively. On the other hand, 1H NMR spectrum
revealed signals at 9.6, 3.86, and 3.84 ppm assignable for
exchangeable NH and two methoxy groups beside the
aromatic and (═CH) signals at their specific regions. The
13C NMR spectrum of 3b showed signals at δ 56.1
and 56.2 assignable for two methoxy groups of the
phenyl ring (Ar–C, C-4, ═CH, and C-3) around
112.0–154.0 ppm and at 170 ppm for C═O of the
pyrazolone moiety.
The N-glucoside derivative 6 was formed by the reaction
of compound
4
with 2,3,4,6-tetra-O-acetyl-α-D-
glucopyranosyl bromide 5 in acetone and aqueous
KOH solution. The IR spectrum of 6 revealed two
characteristic bands at 3157 and 1735 cmꢀ1 attributed to
NH and C═O groups. On the other hand, 1H NMR
spectrum of 6 shows four singlet signals (12 H) around δ
1.96–2.10 ppm assignable for four methyl groups of the
The reaction of 3b with phenylhydrazine in ethanol and
acetic acid solution yielded N-phenylpyrazolo[3,4-c]
Scheme 1. Synthesis of compounds 4–11.
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Synthesis and Anticancer Activity of Some New Fused Pyrazoles
and Their Glycoside Derivatives
acetylated sugar beside the signals corresponding to the
Furthermore, refluxing 3a,b with ethyl acetoacetate
sugar protons in the region δ 3.67–5.72 ppm. The
in the presence of KOH and ethanol gave 3-
0
0
coupling constants of the anomeric protons J1 ,2 equal
9.6 Hz, indicating the N-glycosidic β-configuration,
which agrees with the assigned structure. The 13C NMR
spectrum of the same compound revealed new signals
assignable for (4 CH3) and (4 C═O) groups in addition to
signals for the sugar moiety and the aromatic systems (cf.
Experimental part and Scheme 1).
acetylpyrano[2,3-c]pyrazolone
derivatives
10a,b,
respectively. IR spectra of 10a,b revealed two strong
bands at 1670 and 1600 cmꢀ1 of the two carbonyl groups
1
of the acetylpyranone rings. On the other hand, the H
NMR spectra of compounds 10a,b showed one singlet
signal of the acetyl methyl group at 2.31 ppm and two
doublets at 3.4 and 4.4 ppm for two CH groups
concerning the pyranone ring moiety. The 13C NMR
spectrum of 10a revealed signals of one CH3, 2 (CH),
and 2 (C═O) groups at 28.1, 40.5, 62.5, 165.6, and
200.2 ppm in addition to signals for pyrano-pyrazole
moiety in their specific positions.
Refluxing of 3a,b with thiosemicarbazide in the
presence of absolute EtOH and glacial AcOH afforded
pyrazolo[3,4-c]pyrazole-2(1H)-carbothioamide derivatives
7a,b, respectively. The IR spectra of 7a,b showed bands
1
for NH2 and NH groups. The H NMR spectra of the
same compounds showed new signals of exchangeable
NH2 and NH groups at 6.25 and 9.8 ppm, respectively,
beside signals of the aromatic protons at their specific
regions. The 13C NMR spectra of the same compounds
revealed new signals assignable for C═S at 180 ppm in
addition to signals of the aromatic and bis-pyrazolyl
rings. Compound 7a was reacted with D-xylose in
ethanol and acetic acid solution to afford the sugar
carbothioamide derivative 8. The IR spectrum of 8
showed characteristic bands at 3350–3200 cmꢀ1 for the
OH groups of the xylose moiety and at 3152 cmꢀ1 for
3-Methyl pyrazolo[4,3-f]indazole derivatives 11a,b
were prepared by the reaction of 10a,b with hydrazine
hydrate in absolute EtOH. The IR spectra of 11a,b
revealed the NH band of new pyrazole ring at
3160 cmꢀ1. On the other hand, 1H NMR spectra of
compounds 11a,b showed two singlet signals of
exchangeable NH of the pyrazole and CH of the
pyranone rings at 10.0 and 4.7 ppm in 11a, while in 11b,
1H NMR spectrum showed two NH signals at 10 and
12.57 ppm beside the CH of the pyranone ring at
4.7 ppm. The 13C NMR spectrum of 11a revealed signals
of one CH3 and CH groups at 28.1 and 40.0 ppm in
addition to signals for bis-pyrazolopyran moiety in their
specific positions (cf. Experimental section and Scheme 1).
The reaction of 3a,b with diethylmalonate in
ethanol/piperidine solution yielded 6-oxo-pyrano[2,3-c]
pyrazole-5-carboxylate derivatives 12a,b, respectively.
The IR spectra of 12a,b revealed two strong bands of
two carbonyl groups, one for the ester group and the
other for the carbonyl of the pyranone ring at 1735 and
1550 cmꢀ1, respectively. The 1H NMR spectra of
compounds 12a,b showed signals at δ 1.26 (t) for one
CH3 group and at 4.20 (q) for CH2 group. The 13C NMR
spectra of 12a,b revealed signals of CH3 group and one
CH2 at 15.0 and 61 ppm beside the signals for pyrano-
pyrazole moiety in their specific positions (cf.
Experimental section and Scheme 2).
Refluxing 3a,b with malononitrile in absolute ethanol
and few drops of piperidine gave the 6-aminopyrano[2,3-
c]pyrazole-5-carbonitrile derivatives 13a,b, respectively.
The structures of compounds 13a,b were inferred by their
analytical and spectral data, which are in full agreement
with their proposed structures. The IR spectra of 13a,b
revealed two bands at νmax 3310 and 2230 cmꢀ1
assignable for NH2 and CN groups. On the other hand,
1H NMR spectra of the same compounds showed a signal
at δ 6.74 attributed to exchangeable NH2. The 13C NMR
spectra of 13a and 13b revealed a signal of CN group at
115 ppm beside signals of the pyrano-pyrazole moiety in
their specific positions.
1
NH group of the pyrazole ring. The H NMR spectrum
of 8 showed signals of exchangeable NH group at
9.90 ppm and one singlet signal at 7.52 ppm for (N═CH)
of the condensed sugar beside the signals concerning the
sugar moiety. The 13C NMR spectrum revealed new
signals assignable for the sugar moiety in addition to
signal for C═S, aromatic, and bis-pyrazolyl rings (cf.
Experimental section).
Refluxing 3a,b with hydroxylamine hydrochloride in
absolute EtOH and anhydrous sodium acetate afforded
pyrazolo[3,4-c]isoxazole derivatives 9a,b, respectively.
The IR spectra of 9a,b revealed strong bands of NH
groups of oxazole rings at 3180 and 3170 cmꢀ1 1H
.
NMR spectra of compound 9a showed one singlet
signal of exchangeable NH group at 11.20 ppm of
the oxazole ring beside the other signals in their
1
expected positions, while the H NMR spectrum of 9b
showed two NH signals at 6.59 and 11.20 ppm. The
13C NMR spectra of 9a,b showed new signals
assignable for the new isoxazole moiety beside the
other signals for aromatic and pyrazole rings (cf.
Experimental section).
The reactions of 3a,b with phenylhydrazine,
thiosemicarbazide, or hydroxylamine presumably take
place via the attack of the –NH2 on the carbonyl carbon
in pyrazolones followed by elimination of water to afford
the condensation intermediates X1–X3 (shown in
Scheme 1), which then undergo cyclization by addition
of the NH or OH to the C═C of the chalcone moiety.
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I. F. Nassar, A. F. El Farargy, and F. M. Abdelrazek
Vol 000
Scheme 2. Synthesis of compounds 12–20.
The chalcone derivatives 3a,b were also reacted
with ethyl cyanoacetate in ethanol in presence of
ammonium acetate to afford the pyrazolo[3,4-b]pyridine-
5-carbonitrile derivatives 14a,b, respectively. The IR
spectra of 14a,b revealed bands at νmax 3170, 2220, and
1660 cmꢀ1 for NH, CN, and C═O groups, respectively.
1H NMR spectra of 14a,b showed a signal at 11.10 for
exchangeable NH proton of the pyridone ring in 14a and
two singlet signals at 11.50 and 13.10 ppm for two NH
groups in 14b. The 13C NMR spectrum of 14a,b revealed
signals of CN and C═O groups at 115.0 and
163.0 ppm beside the other signals of aromatic and
pyrazolopyrimidinyl moieties.
methylene to the C═C of the chalcone, enolization of the
carbonyl group to –OH followed by elimination of
ethanol from the esters, or addition of the –OH to a
cyano group of malononitrile to afford pyrano-pyrazoles.
The presence of ammonium acetate transforms the pyran
ring into a pyridine ring.
The reaction of 14a with 2,3,4,6-tetra-O-acetyl-α-D-
glucopyranosyl bromide
5 gave the acetylated O-
glucoside derivative 15. The 1H NMR spectrum of 15
showed the appearance of four singlet signals (12 H) at δ
1.94–2.16 ppm assignable for the four methyl groups
beside the signals corresponding to the sugar protons in
the region δ 3.48–5.20 ppm. The coupling constants of
0
0
The reaction of the chalcones 3a,b with the active
methylene esters or malononitrile (Scheme 2) presumably
takes place via the same mechanism shown in Scheme 1
the anomeric protons J1 ,2 equal 10.0 Hz, indicating the
O-glycosidic β-configuration, which agreed with the
assigned structure. The 13C NMR spectrum of 15
revealed signals at δ 19.32–23.05 ppm for 4 CH3 and at
(X4), involving
a Michael addition of the active
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Synthesis and Anticancer Activity of Some New Fused Pyrazoles
and Their Glycoside Derivatives
167.0–170.0 ppm assignable for (4 C═O) in addition to the
signals for the sugar and the aromatic systems (cf.
Experimental part and Scheme 2).
tetrazolium bromide (MTT) assay. The percentage of the
intact cells was measured and compared with that of the
control and positive control doxorubicin.
Compounds 3a,b were reacted with thiourea in KOH
solution, affording the pyrazolopyrimidine derivatives
16a,b, respectively. The IR spectrum of 16b revealed
characteristic bands at νmax 3200 and 3183 cmꢀ1
The obtained results showed that all compounds showed
dose-dependent anticancer activities.
The IC50 values are shown in Table 1.
From the results obtained in Table 1, we concluded that
four compounds (7a, 8, 18a, and 20a) showed anticancer
IC50 values less than those of the positive control;
this means that these four compounds, compared with
doxorubicin, are good anticancer agents. Two compounds
(17, 20b) showed comparable anticancer activities to
those of the positive control. The rest of the compounds
(9b, 13a, and 13b) showed less anticancer activities than
those of the positive control.
1
attributed to two NH groups. The H NMR spectrum of
16b showed two singlet signals for NH protons at 7.0
and 12.66 ppm, respectively. The 13C NMR spectra of
16b showed the presence of C═S group at 180.7 ppm.
The acetylated thioglycoside derivative 17 was obtained
by the reaction of 16b with 2,3,4,6-tetra-O-acetyl-α-D-
glucopyranosyl bromide 5 in acetone and aqueous KOH
solution. The 1H NMR spectrum of 17 showed the
appearance of four singlet signals (12 H) at
δ
2.01–2.06 ppm assignable for four methyl groups beside
the signals corresponding to the sugar protons in the
region δ 3.67–5.75 ppm. The coupling constant of
EXPERIMENTAL
Chemistry. All melting points were measured using a
Reichert Thermovar apparatus (Reichert Technologies,
Depew, NY) and are uncorrected. Yields listed are of
isolated compounds. The IR spectra were recorded on a
PerkinElmer model 1720 FTIR spectrometer (Perkin-
Elmer, Waltham, MA) for KBr disc. Routine NMR
measurements were made on a Bruker AC-300 or DPX-
300 (Brucker, Elk Grove Village, IL) spectrometer at 300
and 75 MHz respectively. Chemical shifts were reported
in δ scale (ppm) relative to tetramethylsilane as a
reference standard, and the coupling constants J values
are given in Hz. The progress of the reactions was
monitored by thin-layer chromatography using aluminum
silica gel plates 60 F245. Spectral measurements and
elemental analyses were performed at the Micro
Analytical Center at the Faculty of Science, Cairo
University, Cairo, Egypt. Compounds 3a and 16a were
prepared according to literature method [33]: Compound
3a: mp 212–213°C (Lit. mp 215–217°C); 16a: mp 130–
132°C (Lit. mp 136–138°C) [42]. Antitumor activity was
0
0
the anomeric proton J1 ,2 equals 9.2 Hz, indicating the
S-glycosidic β-configuration, which agreed well with
the assigned structure. The 13C NMR spectrum of
17 provided also a confirmation of S-glycosidic β-
configuration of the cyclic sugar.
On the other hand, refluxing 16a,b with hydrazine
hydrate in absolute EtOH yielded the hydrazinyl
derivatives 18a,b, respectively. The IR spectra of 18a,b
showed two bands at νmax 3304–3162 cmꢀ1 for NH2 and
1
NH groups. The H NMR spectra of 18a exhibited two
D2O exchangeable singlet signals at δ 5.91 and 9.1 ppm
assigned to the NH2 and NH groups, while 18b has
two NH signals at 8.10 and 11.10 ppm beside the NH2
signal. The 13C NMR spectrum of 18a revealed the
signals of pyrazolopyrimidine at 105.1–164.6 ppm (cf.
Experimental section and Scheme 2).
When compounds 18a and 18b were refluxed with 4-N,
N-dimethylamino benzaldehyde 19a and/or anthracene-9-
aldehyde 19b, they afforded the Schiff bases 20a,b,
respectively. The IR spectra of 20a,b revealed bands
1
around νmax 3180–3150 cmꢀ1 attributed to NH. The H
NMR spectra of 20a showed the disappearance of the
NH2 singlet signal and the presence of the NH singlet at
9.10 ppm (exchangeable), and N═CH groups are
enveloped in the aromatic region, while those of 20b
revealed two singlet signals (exchangeable) at 8.50 and
10.10 ppm assignable for the two NH protons, beside the
other signals of the N═CH hidden in the aromatic
multiplet. The 13C NMR spectra of the two compounds
provided a further confirmation of the structure (cf.
Experimental section).
Table 1
The anticancer IC50 values of the eight compounds using MTT assay
against the human breast cancer cells.
Compound
MCF-7, IC50 (μM)
7a
8
9b
13a
13b
17
18a
20a
20b
DOXO
10.7
15.4
27.9
23.8
24.3
19.2
14.4
18.5
21.5
18.6
Antitumor activity.
Nine of the synthesized
compounds were examined in vitro for their antitumor
activities against MCF-7 human breast carcinoma cell
line using 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-
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I. F. Nassar, A. F. El Farargy, and F. M. Abdelrazek
Vol 000
evaluated at the Laboratory of Microbiology and Biotech-
H), 7.36 (d, J = 7.2 Hz, 1H, Ar–H), 7.39–7.6 (m, 6H, Ar–
H), 8.50 (s, 1H, NH exchangeable) ppm; 13C NMR
(DMSO-d6): δ 19.32, 19.50, 20.18, 23.05, 56.1, 56.2 (6
CH3), 62.88 (C-60), 64.37 (C-40), 68.65 (C-30), 70.39 (C-
20), 72.82 (C-50), 76 (C4), 95.0 (C-10), 101.6 (C-8),
[111.9–154.6 (Ar–C, C-3 and C-7)], 168.0–170.0 (4
C═O) ppm. Anal. Calcd. for C38H40N4O11 (728.76): C,
62.63; H, 5.53; N, 7.69. Found: C, 62.52; H, 5.60; N, 7.74.
nology at the National Research Center, Giza, Egypt.
4-(3,4-Dimethoxybenzylidene)-5-phenyl-2,4-dihydro-3H-pyrazol-
3-one 3b. A mixture of compound 1b (0.01 mol) and 3,4-
dimethoxybenzaldehyde (0.01 mol) in sodium ethoxide
solution [prepared from sodium metal (0.23 g, 10 mmol)
and 50 mL of absolute ethanol] was refluxed for 10 h, left
to cool, poured on crushed ice, and then neutralized with
diluted HCl. The precipitate that formed was dried and
recrystallized from ethyl acetate to give compound 3b as
a brown powder; yield (75%), mp 175–176°C; IR
(KBr cmꢀ1) νmax: 3045 (CH), 1665 (C═O); 1H NMR
(DMSO-d6): 3.84 (s, 3H, CH3), 3.86 (s, 3H, CH3), 6.6–
6.72 (m, 5H, Ar–H), 7.12 (d, J = 7.2 Hz, 1H, Ar–H), 7.36
(d, J = 7.2 Hz, 1H, Ar–H), 7.39–8.04 (m, 2H, Ar–H, and
═CH), 9.6 (s, 1H, NH exchangeable) ppm; 13C NMR
(DMSO-d6): δ 56.1, 56.2 (2CH3), [112.0–154.0 (Ar–C, C-
4, ═CH, C-3)], 170 (C═O). Anal. Calcd. for C18H16N2O3
(308.34): C, 70.12; H, 5.23; N, 9.09. Found: C, 70.05; H,
5.27; N, 9.0.
General procedure for synthesis of compounds 7a,b.
A
solution of the chalcone 3a or b (0.01 mol) and
thiosemicarbazide (0.01 mol) in ethanol (10 mL) and
glacial acetic acid (2 mL) was refluxed for 5 h, the
reaction mixture was poured on crushed ice and was kept
overnight at room temperature, and the solid that formed
was filtered off, washed with water and dried, and then
recrystallized from methanol to form compounds 7a,b.
3-(4-Chlorophenyl)-4,6-diphenyl-3,6-dihydropyrazolo[3,4-c]
pyrazole-2(1H)-carbothioamide 7a.
Yellow powder; mp
165–167°C; IR (KBr cmꢀ1) νmax: 3304–3162 (NH2 and
1
NH), 1619 (C═N); H NMR (DMSO-d6): δ 5.2 (s, 1H,
3-(3,4-Dimethoxyphenyl)-2,4-diphenyl-1,2,3,6-tetrahydropyrazolo
[3,4-c]pyrazole 4. Compound 3b (0.01 mol) was refluxed
CH pyrazole), 6.20 (s, 1H, NH exchangeable), 7.25 (m,
1H, Ar–H), 7.44–7.86 (m, 9H, Ar–H), 7.89 (d,
J = 6.8 Hz, 2H, Ar–H), 7.99 (d, J = 6.8 Hz, 2H, Ar–H),
9.9 (s, 2H, NH2 exchangeable) ppm; 13C NMR (DMSO-
d6): δ 76 (C4), 101 (C-8), [123–148 (Ar–C, C-3, C-7)],
180 (C═S). Anal. Calcd. for C23H18ClN5S (431.94): C,
63.96, H, 4.20, N, 16.21. Found: C, 64.0; H, 4.30; N,
16.30.
with phenylhydrazine (0.01 mol) in dry ethanol (20 mL)
and glacial acetic acid at 80°C for 8 h. The solvent was
evaporated under reduced pressure. The formed precipitate
was recrystallized from ethanol to produce compound 4 as
a black powder; yield (55%), mp 134–136°C; IR
(KBr cmꢀ1) νmax: 3150 (NH), 1590 (C═N); 1H NMR
(DMSO-d6): 3.84, 3.86 (2s, 6H, 2CH3), 5.10 (s, 1H, CH
pyrazole), 6.6–6.72 (m, 5H, Ar–H), 7.12 (d, J = 7.2 Hz,
1H, Ar–H), 7.36 (d, J = 7.2 Hz, 1H, Ar–H), 7.39–7.6 (m,
6H, Ar–H), 8.50 (s, 1H, NH exchangeable), 9.50 (s, 1H,
NH exchangeable) ppm; 13C NMR (DMSO-d6): δ 56.1,
56.2 (2CH3), 76 (C4), 101 (C-8), [112–154 (Ar–C, C-3,
C-7)]. Anal. Calcd. for C24H22N4O (398.47): C, 72.34; H,
5.57; N, 14.06. Found: C, 72.22; H, 5.60; N, 14.04.
3-(3,4-Dimethoxyphenyl)-4-phenyl-3,6-dihydropyrazolo[3,4-
c]pyrazole-2(1H)-carbothioamide 7b. Brown powder; yield
(50%), mp 190–192°C; IR (KBr cmꢀ1) νmax: 3310–3152
1
(NH2 and NH), 1610 (C═N); H NMR (DMSO-d6): 3.84,
3.86 (2s, 6H, 2CH3), 5.30 (s, 1H, CH pyrazole), 6.25
(bs, 1H, NH), 7.05 (d, J = 7.2 Hz, 1H, Ar–H), 7.36
(d, J = 7.2 Hz, 1H, Ar–H), 7.39–8.04 (m, 6H, Ar–H),
9.56 (s, 2H, NH2 exchangeable), 12.57 (s, 1H, NH
exchangeable); 13C NMR (DMSO-d6): δ 56.1, 56.2
(2CH3), 76 (C4), 101 (C-8), [112–154 (Ar–C, C-3, C-7)],
180 (C═S). Anal. Calcd. for C19H19N5O2S (381.45): C,
59.83; H, 5.02; N, 18.36. Found: C, 59.90; H, 5.10; N,
18.34.
2-[(20,30,40,60-Tetra-O-acetyl-β-D-glucopyranosyl)-6-(4-
(3,4-dimethoxyphenyl)-3,5-diphenyl-5,6-
dihydropyrazolo[3,4-c]pyrazole 6. 2,3,4,6-Tetra-O-acetyl-
α-D-glucopyranosyl bromide 5 (5 mmol) was dissolved in
acetone (20 mL) and added to thione 4 (5 mmol) in
aqueous KOH solution (0.28 g, 5 mmol, 2 mL). The
mixture was stirred at room temperature for 8 h. Then, the
solvent was evaporated under reduced pressure at 40°C,
and the residue was washed with water to remove the
formed potassium bromide. The precipitate formed was
dried and recrystallized from ethanol to yield compound 6
as dark brown powder; yield (60%), mp 104–106°C; IR
(KBr cmꢀ1) νmax: 3157 (NH), 1735 (C═O), 1593 (C═N);
1H NMR (DMSO-d6): δ 1.96–2.1 (4s, 12H, 4 CH3CO),
3.60 (m, 2H, 60,6″-H), 3.84, 3.86 (2s, 6H, 2CH3), 4.10–
4.35 (m, 2H, 50-H, 40H), 5.10 (s, 1H, CH pyrazole), 5.0–
3-(4-Chlorophenyl)-4,6-diphenyl-N-(D-xylotetritolylidene)-3,
6-dihydropyrazolo[3,4-c]pyrazole-2(1H)-carbothioamide 8.
Compound 7a (5 mmol) in ethanol (15 mL) was added to
a well-stirred solution of D-xylose (5 mmol) in water
(5 mL) and glacial acetic acid (1 mL). The mixture was
heated under reflux for 3 h. The solvent was concentrated
and left to cool. The formed precipitate was filtered off,
washed with water, dried, and recrystallized from ethanol
to afford compound 8 as a dark brown powder; yield
(49%), mp 135–1137°C; IR (KBr cmꢀ1) νmax: 3350–3200
1
(OH), 3152 (NH); H NMR (DMSO-d6): δ 3.39–3.59 (m,
5.13 (m, 2H, 20-H, 30H), 5.72 (d, J1 ,2 = 9.6 Hz, 1H, H-
10), 6.6–6.72 (m, 5H, Ar–H), 7.12 (d, J = 7.2 Hz, 1H, Ar–
2H, H-50,500), 3.60 (m, 1H, H-40), 4.20 (m, 1H, H-30),
0
0
4.40 (dd, J = 7.4 Hz, J = 7.8 Hz, 1H, H-20), 4.56 (m, 1H,
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

Month 2018
Synthesis and Anticancer Activity of Some New Fused Pyrazoles
and Their Glycoside Derivatives
OH), 4.84 (d, J = 6.4 Hz, 1H, OH), 5.2 (s, 1H, CH
pyrazole), 5.60 (t, J = 4.6 Hz, 1H, OH), 5.72 (t,
J = 4.6 Hz, 1H, OH), 7.25 (m, 1H, Ar–H), 7.52 (s, 1H,
N═CH), 7.60–7.86 (m, 9H, Ar–H), 7.89 (d, J = 6.8 Hz,
2H, Ar–H), 7.99 (d, J = 6.8 Hz, 2H, Ar–H), 7.52 (d, 1H,
2H, Ar–H) ppm; 13C NMR (DMSO-d6): δ 28.1, 56.1,
56.2 (3CH3), 40.5, 62.5 (2CH), 112.7–156.5
(pyrazolopyranyl-C and Ar–C), 165.6, 200.2 (2C═O)
ppm. Anal. Calcd. for C26H19ClN2O3 (442.90): C, 70.51;
H, 4.32; N, 6.33. Found: C, 70.60; H, 4.30; N, 6.25.
5-Acetyl-4-(3,4-dimethoxyphenyl)-3-phenyl-4,5-dihydropyrano
[2,3-c]pyrazol-6(1H)-one 10b. Black powder; yield (65%),
J1 ,2 = 7.8 Hz, H-10), 9.9 (s, 1H, NH exchangeable) ppm;
13C NMR (DMSO-d6): δ 62.22 (C-50), 63.15 (C-40), 69.32
(C-30), 73.0 (CH), 74.56 (C-20), 121.4–150.03 (pyrazole-C
and Ar–C), 150.44 (C-10), 178.0 (C═S) ppm. Anal. Calcd.
for C28H26ClN5O4S (564.06): C, 59.62; H, 4.65; N,
12.42. Found: C, 59.65; H, 4.50; N, 12.58.
0
0
mp 138–140°C; IR (KBr cmꢀ1) νmax: 3154 (NH), 1670,
1600 (2C═O); 1H NMR (DMSO-d6): δ 2.08 (s, 3H, CH3),
3.52, 4.1 (2d, 2H, 2CH pyranone), 3.84, 3.86 (2s, 6H,
2CH3), 7.05 (d, J = 7.2 Hz, 1H, Ar–H), 7.36 (d,
J = 7.2 Hz, 1H, Ar–H), 7.39–8.04 (m, 6H, Ar–H), 12.50
(s, 1H, NH exchangeable); 13C NMR (DMSO-d6): δ 28.1
(CH3), 40.5, 62.5 (2CH), 112.7–145.5 (pyrazolopyranyl-C
and Ar–C), 165.6, 200.2 (2C═O) ppm. Anal. Calcd. for
C22H20N2O5 (392.41): C, 67.34; H, 5.14; N, 7.14. Found:
C, 67.40; H, 5.20; N, 7.10.
General procedure for synthesis of compounds 9a,b.
A
mixture of 3a,b (0.01 mol) and hydroxylamine
hydrochloride (0.01 mol) in absolute ethanol (30 mL) and
anhydrous sodium acetate (0.01 mol) was refluxed for
10 h, the mixture was poured on crushed ice, and the
solid formed was filtered off, washed with water and
dried, and then recrystallized from methanol to form
compounds 9a,b.
General procedure for synthesis of compounds 11a,b.
A
mixture of compounds 10a,b (0.01 mol) and hydrazine
hydrate (0.015 mol) in ethanol (20 mL) was heated for
6 h. The product was allowed to cool at room
temperature. The solvent was evaporated under vacuum.
The formed precipitate was recrystallized from ethyl
alcohol to produce compounds 11a,b.
3-(4-Chlorophenyl)-4,6-diphenyl-3,6-dihydro-1H-pyrazolo[3,
4-c]isoxazole 9a.
Yellow powder; mp 165–167°C;
IR (KBr cmꢀ1) νmax: 3180 (NH); H NMR (DMSO-d6):
δ 7.38 (m, 1H, Ar–H), 7.49–7.68 (m, 9H, Ar–H), 7.92
(d, J = 6.8 Hz, 2H, Ar–H), 7.95 (d, J = 6.8 Hz, 2H,
Ar–H), 11.20 (s, 1H, NH exchangeable) ppm; 13C
NMR (DMSO-d6): δ 87 (C4), 101 (C-8), [123–148
(Ar–C, C-3, C-7)]. Anal. Calcd. for C22H16ClN3O
(373.84): C, 70.68; H, 4.31; N, 11.24. Found: C, 70.70;
1
4-(4-Chlorophenyl)-5-methyl-1,3-diphenyl-1,4,7,8-tetrahydro
pyrazolo[4,3-f]indazole 11a.
Grey powder; mp 163–
165°C; IR (KBr cmꢀ1) νmax: 3160 (NH); 1H NMR
(DMSO-d6): δ 1.9 (s, 3H, CH3), 4.46 (s, 1H, CH pyran),
7.2 (m, 1H, Ar–H), 7.49–7.88 (m, 9H, Ar–H), 7.92 (d,
J = 6.8 Hz, 2H, Ar–H), 7.96 (d, J = 6.8 Hz, 2H, Ar–H),
10.0 (s, 1H, NH exchangeable) ppm; 13C NMR (DMSO-
d6): δ 28.1 (CH3), 41.0 (CH), [112.7–145.5 (bis-pyrazolo-
pyran-C and Ar–C)] ppm. Anal. Calcd. for C26H19ClN4O
(438.92): C, 71.15; H, 4.36; N, 12.77. Found: C, 71.0; H,
4.35; N, 12.80.
H, 4.30; N, 11.15.
3-(3,4-Dimethoxyphenyl)-4-phenyl-3,6-dihydro-1H-pyrazolo
[3,4-c]isoxazole 9b. Bale brown powder; yield (60%), mp
1
185–187°C; IR (KBr cmꢀ1) νmax: 3170 (NH); H NMR
(DMSO-d6): 3.84, 3.86 (2s, 6H, 2CH3), 5.90 (s, 1H, CH
pyrazole), 7.05 (d, J = 7.2 Hz, 1H, Ar–H), 7.36 (d,
J = 7.2 Hz, 1H, Ar–H), 7.39–8.04 (m, 6H, Ar–H), 6.59
(s, 1H, NH exchangeable), 11.20 (s, 1H, NH
exchangeable); 13C NMR (DMSO-d6): δ 56.1, 56.2
(2CH3), 87 (C4), 101 (C-8), [112–154 (Ar–C, C-3, C-7)].
Anal. Calcd. for C18H17N3O3 (323.35): C, 66.86; H, 5.30;
N, 13.00. Found: C, 66.90; H, 5.40; N, 13.04.
4-(3,4-Dimethoxyphenyl)-3-methyl-5-phenyl-4,7-dihydro-1H-
pyrano[2,3-c:6,5-c0]dipyrazole 11b.
Brown powder; yield
(45%), mp 120–122°C; IR (KBr cmꢀ1) νmax: 3200, 3160
1
(2NH); H NMR (DMSO-d6): δ 1.9 (s, 3H, CH3), 4.7 (s,
1H, CH pyranone), 3.84, 3.86 (2s, 6H, 2CH3), 7.05 (d,
J = 7.2 Hz, 1H, Ar–H), 7.36 (d, J = 7.2 Hz, 1H, Ar–H),
7.39–8.04 (m, 6H, Ar–H), 10.0 (s, 1H, NH
exchangeable), 12.57 (s, 1H, NH exchangeable); 13C
NMR (DMSO-d6): δ 28.1, 56.1, 56.2 (3CH3), 41.0 (CH),
[112.7–156.5 (bis-pyrazolo-pyran-C and Ar–C)] ppm.
Anal. Calcd. for C22H20N4O3 (388.43): C, 68.03; H, 5.19;
N, 14.42. Found: C, 68.10; H, 5.20; N, 14.34.
General procedure for synthesis of compounds 10a,b.
A
solution of 3a,b (0.01 mol), ethyl acetoacetate (0.01 mol),
and ammonium acetate (0.01 mol) in absolute ethanol
(15 mL) containing aqueous KOH solution (1 mL, 10%)
was refluxed for 2 h and left overnight at room
temperature. The precipitate formed was filtered off and
recrystallized from ethyl acetate to produce compounds
10a,b.
General procedure for synthesis of compounds 12a,b.
A
solution of compounds 3a,b (0.01 mol) and
diethylmalonate (0.01 mol) in absolute ethanol (30 mL)
and few drops of piperidine was refluxed for 6 h. The
product was cooled at room temperature. Excess solvent
was evaporated under vacuum. The precipitate was
recrystallized from methanol to produce compounds 12a,b.
5-Acetyl-4-(4-chlorophenyl)-1,3-diphenyl-4,5-dihydropyrano
[2,3-c]pyrazol-6(1H)-one 10a.
Yellow powder; mp
1
120–122°C; H NMR (DMSO-d6): IR (KBr cmꢀ1) νmax
:
1670, 1600 (2C═O); δ 2.31 (s, 3H, CH3), 3.4, 4.4 (2d,
2H, 2CH), 7.38 (m, 1H, Ar–H), 7.49–7.68 (m, 9H, Ar–
H), 7.89 (d, J = 6.8 Hz, 2H, Ar–H), 7.99 (d, J = 6.8 Hz,
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

I. F. Nassar, A. F. El Farargy, and F. M. Abdelrazek
Vol 000
Ethyl 4-(4-chlorophenyl)-6-oxo-1,3-diphenyl-1,6-dihydropyrano
[2,3-c]pyrazole-5-carboxylate 12a.
General procedure for synthesis of compounds 14a,b.
A
solution of compound 3a or 3b (0.01 mol) and ethyl
cyanoacetate (0.01 mol) and ammonium acetate
(10 mmol) in ethanol (30 mL) was refluxed for 9 h and
allowed to cool at room temperature. The precipitate
formed after evaporation of excess solvent was
recrystallized from ethanol to produce compounds 14a,b.
Brown powder; mp
120–122°C; IR (KBr cmꢀ1) νmax: 1735, 1550 (2C═O);
1H NMR (DMSO-d6): δ 1.26 (t, J = 13.2 Hz, 3H, CH3),
4.20 (q, J = 13.2 Hz, 2H, CH2), δ 7.22–7.26 (m, 1H,
Ar–H), 7.28–7.61 (m, 9H, Ar–H), 7.9 (d, J = 6.8 Hz, 2H,
Ar–H), 8.10 (d, J = 6.8 Hz, 2H, Ar–H); 13C NMR
(DMSO-d6): δ 15.0 (CH3), 61.0 (CH2), 112.0 (C-9),
[118.5–153.0 (Ar–C, C-3, C-8, C-4, C-5)], 159, 163
(2C═O) ppm. Anal. Calcd. for C27H19ClN2O4 (470.91):
C, 68.87; H, 4.07; N, 5.95. Found: C, 70.0; H, 4.05;
4-(4-Chlorophenyl)-6-oxo-1,3-diphenyl-6,7-dihydro-1H-pyrazolo
[3,4-d]pyridine-5-carbonitrile 14a.
Yellow powder; yield
(55%), mp 240–242°C; IR (KBr cmꢀ1) νmax: 3170 (NH),
1
2320 (CN), 1660 (C═O); H NMR (DMSO-d6): δ 7.38
N, 6.05.
(m, 1H, Ar–H), 7.49–7.68 (m, 9H, Ar–H), 7.92
(d, J = 6.8 Hz, 2H, Ar–H), 7.95 (d, J = 6.8 Hz, 2H,
Ar–H), 11.10 (s, 1H, NH exchangeable) ppm; 13C
NMR (DMSO-d6): δ 95 (pyridine-CH), 100 (C-9),
115.0 (CN), [118.0–148.0 (Ar–C, C-3, C-8, C-4)], 163
(C═O) ppm. Anal. Calcd. for C25H15ClN4O (422.87):
C, 71.01; H, 3.58; N, 13.25. Found: C, 70.90; H, 3.50;
N, 13.15.
Ethyl 4-(3,4-dimethoxyphenyl)-6-oxo-3-phenyl-1,6-dihydro
pyrano[2,3-c]pyrazole-5-carboxylate 12b.
Brown powder;
yield (56%), mp 123–125°C; IR (KBr cmꢀ1) νmax: 3170
1
(NH), 1730, 1665 (2C═O); H NMR (DMSO-d6): δ 1.26
(t, J = 13.2 Hz, 3H, CH3), 3.84, 3.86 (2s, 6H, 2CH3),
4.20 (q, J = 13.2 Hz, 2H, CH2), 7.05 (d, J = 7.2 Hz, 1H,
Ar–H), 7.36 (d, J = 7.2 Hz, 1H, Ar–H), 7.39–8.04 (m,
6H, Ar–H), 13.10 (s, 1H, NH exchangeable) ppm; 13C
NMR (DMSO-d6): δ 15.0, 56.1, 56.2 (3CH3), 61.0
(CH2), 105.0 (C-9), [108.5–153.0 (Ar–C, C-3, C-8, C-4,
C-5)], 160, 163 (2C═O) ppm. Anal. Calcd. for
C23H20N2O6 (362.39): C, 65.71; H, 4.80; N, 6.66. Found:
C, 65.70; H, 4.92; N, 6.45.
4-(3,4-Dimethoxyphenyl)-6-oxo-3-phenyl-6,7-dihydro-1H-
pyrazolo[3,4-b]pyridine-5-carbonitrile 14b. Yellow powder;
yield (55%), mp 138–140°C; IR (KBr cmꢀ1) νmax: 3200,
1
3170 (2NH), 2300 (CN), 1660 (C═O); H NMR (DMSO-
d6): δ 3.84, 3.86 (2s, 6H, 2CH3), 7.05 (d, J = 7.2 Hz, 1H,
Ar–H), 7.36 (d, J = 7.2 Hz, 1H, Ar–H), 7.37–7.64 (m,
6H, Ar–H), 11.50 (s, 1H, NH exchangeable), 13.10 (s,
1H, NH exchangeable); 13C NMR (DMSO-d6): δ 56.1,
56.2 (2CH3), 95 (pyridine-CH), 100 (C-9), 115.0 (CN),
[118.0–148.0 (Ar–C, C-3, C-8, C-4)], 163 (C═O) ppm.
Anal. Calcd. for C21H16N4O3 (372.38): C, 67.73; H, 4.33;
N, 15.05. Found: C, 67.80; H, 4.20; N, 15.10.
General procedure for synthesis of compounds 13a,b.
A
mixture of 3a,b (0.01 mol) and malononitrile (0.01 mol) in
absolute ethanol (30 mL) was refluxed for 4 h. The product
was allowed to cool at room temperature. Excess ethanol
was evaporated under vacuum. The precipitate was
recrystallized from ethyl acetate to produce compounds
13a,b.
6-(4-(4-Chlorophenyl)-5-cyano-1,3-diphenyl-6-[(20,30,40,60-
tetra-O-acetyl-β-D-glucopyranosyl)-1H-pyrazolo[3,4-d]pyridine
15. 2,3,4,6-Tetra-O-acetyl-α-D-glucopyranosyl bromide
6-Amino-4-(4-chlorophenyl)-6-oxo-1,3-diphenyl-1,6-dihydro
pyrano[2,3-c]pyrazole-5-carbonitrile 13a.
Black powder;
mp 175–177°C; IR (KBr cmꢀ1) νmax: 3310 (NH2), 2300
(CN); 1H NMR (DMSO-d6): δ 6.74 (bs, 2H, NH2
exchangeable), 7.24–7.26 (m, 1H, Ar–H), 7.28–7.61 (m,
9H, Ar–H), 7.9 (d, J = 6.8 Hz, 2H, Ar–H), 8.10 (d,
J = 6.8 Hz, 2H, Ar–H); 13C NMR (DMSO-d6): δ 25
(pyran-CH), 59, 115.0 (CN), [119.5–173.0 (Ar–C, C-3,
C-8, C-4, C-6)]. Anal. Calcd. for C25H17ClN4O (424.89):
C, 70.67; H, 4.03; N, 13.19. Found: C, 70.70; H, 4.05;
5 (5 mmol) dissolved in acetone (20 mL) was added to a
mixture of compound 14a (5 mmol) and aqueous
potassium hydroxide (0.28 g, 5 mmol, 2 mL). The
reaction mixture was stirred at room temperature for 6 h.
The solvent was evaporated under reduced pressure at
40°C, and the residue was washed with distilled water to
remove the formed potassium bromide. The product was
dried and recrystallized from ethanol to yield a mixture
of compound 15. Dark brown powder; yield (54%), mp
126–128°C; IR (KBr cmꢀ1) νmax: 1735 (C═O), 1593
N, 13.25.
6-Amino-4-(3,4-dimethoxyphenyl)-6-oxo-1,3-diphenyl-1,6-
dihydropyrano[2,3-c]pyrazole-5-carbonitrile 13b.
Black
powder; yield (66%), mp 135–137°C; IR (KBr cmꢀ1
)
(C═N); H NMR (DMSO-d6): δ 1.94–2.16 (4s, 12H, 4
1
1
νmax: 3310–3150 (NH2 and NH), 2300 (CN); H NMR
CH3CO), 3.48 (m, 2H, 60,6″-H), 3.8–3.88 (m, 2H, 50-H,
40H), 4.3–4.39 (m, 2H, 20-H, 30H), 5.2 (d,
(DMSO-d6): δ 3.84, 3.86 (2s, 6H, 2CH3), 6.74 (bs, 2H,
NH2 exchangeable), 7.05 (d, J = 7.2 Hz, 1H, Ar–H), 7.36
(d, J = 7.3 Hz, 1H, Ar–H), 7.39–8.04 (m, 6H, Ar–H),
13.10 (s, 1H, NH exchangeable) ppm; 13C NMR (DMSO-
d6): δ 25 (pyran-CH), 56.1, 56.2 (2CH3), 63, 115.0 (CN),
[119.5–173.0 (Ar–C, C-3, C-8, C-4, C-6)]. Anal. Calcd.
for C21H18N4O3 (374.40): C, 67.37; H, 4.85; N, 14.96.
Found: C, 67.10; H, 4.95; N, 15.0.
J1 ,2 = 10.0 Hz, 1H, H-10), δ 7.22–7.26 (m, 1H, Ar–H),
7.28–7.61 (m, 9H, Ar–H), 7.9 (d, J = 6.8 Hz, 2H,
0
0
Ar–H), 8.03 (d,
J =
6.8 Hz, 2H, Ar–H); 13C
NMR (DMSO-d6): δ 19.32, 19.50, 20.18, 23.05 (4 CH3),
62.88 (C-60), 64.37 (C-40), 68.65 (C-30), 70.39 (C-20),
72.82 (C-50), 88.95 (C-10), 101.6 (C-9), [118.9–158.6
(Ar–C, C-3, C-4 and C-8)], 167.0–170.0 (4 C═O), 172.1
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

Month 2018
Synthesis and Anticancer Activity of Some New Fused Pyrazoles
and Their Glycoside Derivatives
(C-6) ppm. Anal. Calcd. for C39H33ClN4O10 (753.16):
C, 62.20; H, 4.42; N, 7.44. Found: C, 62.40; H, 4.40;
N, 7.50.
4-(4-Chlorophenyl)-6-hydrazinyl-1,3-diphenyl-1H-pyrazolo[3,
4-d]pyrimidine 18a.
Brown powder; yield (55%), mp
126–128°C; IR (KBr cmꢀ1) νmax: 3304–3162 (NH2 and
1
NH), 1619 (C═N); H NMR (DMSO-d6): δ 5.91 (bs, 2H,
4-(3,4-Dimethoxyphenyl)-3-phenyl-1H-pyrazolo[3,4-d]pyrimidin-
NH2 exchangeable), δ 7.24–7.26 (m, 1H, Ar–H), 7.28–
7.61 (m, 9H, Ar–H), 7.9 (d, J = 6.8 Hz, 2H, Ar–H), 8.10
(d, J = 6.8 Hz, 2H, Ar–H); 9.10 (s, 1H, NH exchangeable)
ppm; 13C NMR (DMSO-d6): δ 105.0 (C-9), [123.9–164.6
(Ar–C, C-3, C-8, C-6, and C-4)] ppm. Anal. Calcd. for
C23H17ClN6 (412.88): C, 66.91; H, 4.15; N, 20.36. Found:
6(7H)-thione 16b.
Thiourea (0.01 mmol) was added to
compound 3b (0.01 mol) and dissolved in sodium ethoxide
solution [prepared from sodium metal (0.23 g, 10 mmol) in
50 mL absolute ethanol]. The reaction mixture was
refluxed for 12 h. The mixture was left to cool, poured on
crushed ice, and neutralized with diluted hydrochloric acid.
The precipitate was collected by filtration, dried, and
recrystallized from ethanol to give compound 16b. Yellow
C, 66.90; H, 4.05; N, 20.35.
4-(3,4-Dimethoxyphenyl)-6-hydrazinyl-3-phenyl-1H-pyrazolo
powder; yield (65%), mp 156–158°C; IR (KBr cmꢀ1) νmax
:
[3,4-d]pyrimidine 18b.
Brown powder; yield (70%), mp
115–116°C; IR (KBr cmꢀ1) νmax: 3300–3160 (NH2 and
NH), 1619 (C═N); 1H NMR (DMSO-d6): δ 3.84, 3.86 (2s,
6H, 2CH3), 6.1 (bs, 2H, NH2 exchangeable), 7.05 (d, 1H,
J = 7.2 Hz, Ar–H), 7.36 (d, 1H, J = 7.2 Hz, Ar–H), 7.39–
8.04 (m, 6H, Ar–H), 8.10 (s, 1H, NH exchangeable), 11.10
(s, 1H, NH exchangeable) ppm; 13C NMR (DMSO-d6): δ
56.1, 56.2 (2CH3), 105.0 (C-9), [108.5–163.0 (Ar–C, C-3,
C-8, C-4, C-6)]. Anal. Calcd. for C19H18N6O2 (362.39): C,
62.97; H, 5.01; N, 23.19. Found: C, 63.10; H, 5.02; N, 23.25.
3200, 3183 (2NH); 1H NMR (DMSO-d6): δ 3.83, 3.86 (2s,
6H, 2CH3), 7.05 (d, J = 7.2 Hz, 1H, Ar–H), 7.36 (d,
J = 7.2 Hz, 1H, Ar–H), 7.39–8.04 (m, 5H, Ar–H), 7.48 (s,
1H, Ar–H), 7.0 (s, 1H, NH exchangeable), 12.66 (s, 1H,
NH exchangeable) ppm; 13C NMR (DMSO-d6): δ 56.1,
56.2
(2CH3),
[105.0–164.0
(Ar–C
and
pyrazolopyrmidinyl-C)], 180.7 (C═S) ppm. Anal. Calcd.
for C19H16N4O2S (364.42): C, 62.62; H, 4.43; N, 15.37.
Found: C, 62.55; H, 4.57; N, 15.49.
4-(3,4-Dimethoxyphenyl)-3-phenyl-1-phenyl-6-[(2,3,4,6-tetra-
O-acetyl-β-D-glucopyranosyl)thio]-1H-pyrazolo[3,4-d]pyrimidine
17. 2,3,4,6-Tetra-O-acetyl-α-D-glucopyranosyl bromide 5
(5 mmol) was dissolved in acetone (20 mL) and added to
thione 16b (5 mmol) in aqueous KOH solution (0.28 g,
5 mmol, 2 mL). The reaction mixture was stirred at room
temperature for 8 h. The solvent was evaporated
under reduced pressure at 40°C, and the residue was
washed with distilled water to remove the formed
potassium bromide. The obtained product was dried and
recrystallized from ethanol to yield compound 17. Dark
brown powder; yield (45%), mp 115–117°C; IR
(KBr cmꢀ1) νmax: 3173 (NH), 1730 (CO); 1H NMR
(DMSO-d6): δ 2.01–2.06 (4s, 12H, 4 CH3CO), 3.83 and
3.86 (2s, 6H, 2OCH3), 3.67 (m, 2H, 60,6″-H), 4.10–4.25
(m, 2H, 50-H, 40H), 5.05–5.10 (m, 2H, 20-H, 30H), 5.75 (d,
General procedure for synthesis of compounds 20a,b.
A
solution of compounds 18a,b (0.01 mol) and aromatic
aldehydes as 4-N,N-dimethylamino benzaldehyde 19a or
anthracene-9-aldehyde 19b in absolute ethanol (30 mL)
and glacial acetic acid (one to two drops) was refluxed
for 10 h. The solid product was allowed to cool at room
temperature. The solvent was evaporated under vacuum.
The powder was recrystallized from ethyl alcohol to
produce compounds 20a,b.
4-(2-(4-(4-Chlorophenyl)-1,3-diphenyl-1H-pyrazolo[3,4-d]
pyrimidin-6yl)hydrazono)methyl-N,N-dimethylaniline 20a.
Grey powder; mp 160–162°C; IR (KBr cmꢀ1) νmax
:
3180–3150 (NH), 3050 (CH); 1H NMR (DMSO-d6): δ
2.99, 3.01 (2s, 6H, 2CH3), 7.1 (d, J = 6.6 Hz, 2H, Ar–H),
7.23 (s, 1H, N═CH), 7.28–7.61 (m, 10H, Ar–H, and
N═CH), 7.74 (d, J = 6.6 Hz, 2H, Ar–H), 7.9 (d,
J = 6.8 Hz, 2H, Ar–H), 8.10 (d, J = 6.8 Hz, 2H, Ar–H);
9.10 (s, 1H, NH exchangeable) ppm; 13C NMR
(DMSO-d6): δ 40.1, 40.2 (2CH3), 105.0 (C-9), [112.0–
164.6 (Ar–C, N═CH, C-3, C-8, C-6, and C-4)] ppm. Anal.
Calcd. for C32H26ClN7 (544.06): C, 70.65; H, 4.82; N,
J1 ,2 = 9.2 Hz, 1H, H-10), 7.05 (d, 1H, J = 7.2 Hz, Ar–H),
7.36 (d, 1H, J = 7.2 Hz, Ar–H), 7.39–8.04 (m, 6H, Ar–
H), 11.1 (s, 1H, NH exchangeable) ppm; 13C NMR
(DMSO-d6): δ 19.32, 19.50, 20.18, 20.27, 56.10, 56.20 (6
CH3), 62.88 (C-60), 64.37 (C-40), 68.65 (C-30), 70.39 (C-
20), 72.82 (C-50), 88.95 (C-10), 105.0 (C-9), [108.5–163
(Ar–C, C-3, C-4, C-8)], 167.0–170.0 (4 C═O), 172.2 (C-
6) ppm. Anal. Calcd. for C33H34N4O11S (694.71): C,
57.05; H, 4.93; N, 8.06. Found: C, 57.25; H, 4.90; N, 8.0.
0
0
18.02. Found: C, 70.70; H, 4.95; N, 18.05.
6-(2-Anthracene-9-ylmethylene)hydrazinyl)-4-(3,4-dimethoxy
phenyl)-3-phenyl-1H-pyrazolo[3,4-d]pyrimidine 20b.
Dark
brown powder; yield (50%), mp 150–152°C; IR
(KBr cmꢀ1) νmax: 3210, 3180 (2NH), 3052 (CH); 1H NMR
(DMSO-d6): δ 3.84, 3.86 (2s, 6H, 2CH3), 7.07 (s, 1H, Ar–
H), 7.25–7.40 (d, J = 6.8 Hz, 2H, Ar–H), 7.42–7.75 (m,
12H, Ar–H + CH methine), 7.90 (d, J = 6.8 Hz, 2H, Ar–
H), 8.0 (s, 1H, Ar–H), 8.50 (s, 1H, NH exchangeable),
10.10 (s, 1H, NH exchangeable) ppm; 13C NMR (DMSO-
d6): δ 56.26, 56.5 (2CH3), 102.66 (C-9), [120.3–172.48
(Ar–C, N═CH, C-3, C-8, C-4, C-6)]. Anal. Calcd. for
General procedure for synthesis of compounds 18a,b.
A
solution of compounds 16a,b (0.01 mol) and hydrazine
hydrate (0.15 mol) in absolute ethanol (30 mL) was
refluxed for 6 h. The mixture was allowed to cool at
room temperature. The solvent was evaporated under
vacuum. The formed precipitate was recrystallized from
ethyl acetate to produce compounds 18a,b.
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

I. F. Nassar, A. F. El Farargy, and F. M. Abdelrazek
Vol 000
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Malkowitz, L.; Springer, M. S.; Gould, S. L.; DeMartino, J. A.; Siciliano,
S. J.; Kwei, G. Y.; Carella, A.; Carver, G.; Holmes, K.; Schleif, W. A.;
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V. D.; Corelli, F. J Med Chem 2008, 51, 1560.
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2008, 51, 142.
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H.-S.; Pawlitz, J. L.; Williams, J. M.; Stallings, W. C.; Geng, L.;
Naraian, A. S.; Koszyk, F. J.; Stealey, M. A.; Xu, S. D.; Weier, R. M.;
Hanson, G. J.; Mourey, R. J.; Compton, R. P.; Mannich, S. J.;
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A.; Mosti, L. Eur J Med Chem 2008, 43, 2627.
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Amparo, E.; Wells, B.; Ellis, C.; He, M. Y.; Alexander, R. S.; Rossi, K.
A.; Smallwood, A.; Wong, P. C.; Luettgen, J. M.; Rendina, A. R.; Knabb,
R. M.; Mersinger, L.; Kettner, C.; Bai, S.; He, K.; Wexler, R. R.; Lam, P.
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C34H26N6O2 (550.62): C, 74.17; H, 4.76; N, 15.26. Found:
C, 74.10; H, 4.80; N, 15.25.
In vitro anticancer activity.
The antitumor activity
against the human breast cancer cells was assessed using
the MTT assay [43–45]. This cancer cell line was
purchased from ATCC (Rockville, MD, USA).
The cells were cultured in a 96-well sterile microplate
(5 × 104 cells per well) at 37°C in Roswell Park Memorial
Institute medium (RPMI-1640) supplemented with 10%
heat-inactivated fetal bovine serum and 100 U/mL of both
penicillin and streptomycin in a 5% CO2-humidified
atmosphere. After 24 h, the media were removed and a
fresh serum-free medium (90 μL/well) was added together
with 10 μL of a series of compounds or doxorubicin
(positive control) concentrations in DMSO for 48 h. Then,
media were removed, and MTT (40 μL of 2.5 mg/mL)
was added to each well and incubated for 4 h; 200 μL of
DMSO was added to solubilize the formazan dye crystals
(purple color). With the use of a SpectraMax® Paradigm®
Multi-Mode Microplate Reader, the absorbance was
measured at 590 nm. Each experiment was repeated on
three different days and conducted in triplicate. The
relative cell cytotoxicity was measured according to the
following equation:
%cytotoxicity ¼ ð1 ꢀ As=AbÞꢁ 100;
where As is the absorbance of each sample and Ab the ab-
sorbance of the blank. The probit analysis using the SPSS
software program (version 20, SPSS Inc., Chicago, IL,
USA) was used to determine each IC50.
[19] Li, Y.-L.; Fevig, J. M.; Cacciola, J.; Buriak, J.; Rossi, K. A.;
Jona, J.; Knabb, R. M.; Luettgen, J. M.; Wong, P. C.; Bai, S. A.; Wexler,
R. R.; Lam, P. Y. S. Bioorg Med Chem Lett 2006, 16, 5176.
[20] Fevig, J. M.; Cacciola, J.; Buriak, J.; Rossi, K. A.; Knabb, R.
M.; Luettgen, J. M.; Wong, P. C.; Bai, S. A.; Wexler, R. R.; Lam, P. Y.
S. Bioorg Med Chem Lett 2006, 16, 3755.
[21] Pinto, D. J. P.; Orwat, M. J.; Koch, S.; Rossi, K. A.; Alexander, R.
S.; Smallwood, A.; Wong, P. C.; Rendina, A. R.; Luettgen, J. M.; Knabb, R.
M.; He, K.; Xin, B.; Wexler, R. R.; Lam, P. Y. S. J Med Chem 2007, 50, 5339.
[22] Chou, L.-C.; Huang, L.-J.; Yang, J.-S.; Lee, F.-Y.; Teng, C.-M.;
Kuo, S.-C. Bioorg Med Chem 2007, 15, 1732.
[23] Nassar, I. F.; El Farargy, A. F.; Abdelrazek, F. M.; Ismail, N. S.
M. Nucleosides Nucleotides Nucleic Acids 2017, 36, 275.
[24] Peat, A. J.; Boucheron, J. A.; Dickerson, S. H.; Garrido, D.;
Mills, W.; Peckham, J.; Preugschat, F.; Smalley, T.; Schweiker, S. L.;
Wilson, J. R.; Wang, T. Y.; Zhou, H. Q.; Thomson, S. A. Bioorg Med
Chem Lett 2004, 14, 2121.
Acknowledgments. F.M.A. thanks the Alexander von Humboldt
Foundation (Germany) for the continuous help and support
through granting short research fellowships. Thanks are also due
to Professor Peter Metz, Institute für Organische Chemie, TU
Dresden, for his repeated invitations and generous hospitality.
A.F.E.F. thanks Professor Norbert Krause, Lehrstuhl für
Organische Chemie, TU Dortmund, for his repeated invitations,
kind hospitality, and the facilities offered.
[25] Rashad, A. E.; Hegab, M. I.; Abdel-Megeid, R. E.; Micky, J.
A.; Abdel-Megeid, F. M. E. Bioorg Med Chem 2008, 16, 7102.
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[29] Abdelrazek, F. M.; Fadda, A. A.; Elsayed, A. N. Synth
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Synthesis and Anticancer Activity of Some New Fused Pyrazoles
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