Synthesis and anticancer activity of novel 2-pyridyl hexahyrocyclooctathieno[2,3-d]pyrimidine derivatives

Article abstract of DOI:10.1016/j.ejmech.2013.02.011

A series of new 2-pyridyl hexahydrocycloocta [4,5]thieno[2,3-d]pyrimidines with different substituents as C-4 position was synthesized. The anticancer activity of the newly synthesized compounds was tested in vitro using a two-stage process utilizing 60 different human tumor cell lines representing leukemia, melanoma and cancers of lung, colon, central nervous system, ovary, kidney, prostate as well as breast. Compounds 4a, 6a, 7a, 7d and 7g showed potent anticancer activity at low concentrations against most of the used human tumor cell lines comparable with doxorubicin as standard potent anticancer drug (average log₁₀ GI₅₀ over all cell lines = -6.85). Also, compound 4b was selective against SNB-75 (CNS cancer) log₁₀ GI ₅₀ = -5.57. Interestingly, compound 7e exhibited promising selectivity against 13 tumor cell lines showing growth inhibition percentages between 54.05 and 89.23.

Full text of DOI:10.1016/j.ejmech.2013.02.011

European Journal of Medicinal Chemistry 63 (2013) 224e230
Contents lists available at SciVerse ScienceDirect
European Journal of Medicinal Chemistry
journal homepage: http://www.elsevier.com/locate/ejmech
Original article
Synthesis and anticancer activity of novel 2-pyridyl hexahyrocyclooctathieno
[2,3-d]pyrimidine derivatives
*
Asmaa E. Kassab , Ehab M. Gedawy
Pharmaceutical Organic Chemistry Department, Cairo University, 33 Kasr El-Aini Street, Cairo 11562, Egypt
a r t i c l e i n f o
a b s t r a c t
Article history:
A series of new 2-pyridyl hexahydrocycloocta [4,5]thieno[2,3-d]pyrimidines with different substituents
as C-4 position was synthesized. The anticancer activity of the newly synthesized compounds was tested
in vitro using a two-stage process utilizing 60 different human tumor cell lines representing leukemia,
melanoma and cancers of lung, colon, central nervous system, ovary, kidney, prostate as well as breast.
Compounds 4a, 6a, 7a, 7d and 7g showed potent anticancer activity at low concentrations against most
of the used human tumor cell lines comparable with doxorubicin as standard potent anticancer drug
(average log10 GI50 over all cell lines ¼ ꢀ6.85). Also, compound 4b was selective against SNB-75 (CNS
cancer) log10 GI50 ¼ ꢀ5.57. Interestingly, compound 7e exhibited promising selectivity against 13 tumor
cell lines showing growth inhibition percentages between 54.05 and 89.23.
Received 5 October 2012
Received in revised form
8 February 2013
Accepted 12 February 2013
Available online 19 February 2013
Keywords:
2-Pyridyl hexahydrocyclooctathieno[2,3-d]
pyrimidines
2-Pyridyl hexahydrocyclooctathieno[2,3-d]
pyrimidin-4(3H)-ones
Synthesis
Ó 2013 Elsevier Masson SAS. All rights reserved.
Anticancer activity
1. Introduction
be screened as anticancer agent, we reported in previous works the
synthesis of new hexahydrocycloocta [4,5]thieno[2,3-d]pyrimi-
Cancer remains as a major cause of death worldwide so there is
an ongoing need for discovery and development of effective anti-
cancer agents. Recently, the thieno[2,3-d]pyrimidine core was
evaluated as bioisostere of 4-anilinoquinazoline core which in-
cludes potent marketed anticancer drugs for example gefitinib
(IressaÔ) [1], erlotinib (TarcevaÔ) [2] and tandutinib (MLN518)
(phase II clinical trials) [3] (Fig. 1). A large number of thieno[2,3-d]
pyrimidine derivatives were found to be active against different
cancer types exerting their antitumor activities via different
mechanisms [4e23]. Consequently, the thieno[2,3-d]pyrimidine
ring system constitutes an attractive target for the design of new
anticancer drugs through wide structure variations. A research
group reported that cycloalkyl ring fused to thiophene was essen-
tial for the anticancer activity of thieno[2,3-d]pyrimidines [7]. Then
several studies support the anticancer activity of thieno[2,3-d]py-
rimidines fused with five-, six- or seven-membered cycloalkyl
lipophilic moieties [10,15e18]. Because 8-membered cycloalkyl ring
was not incorporated in the synthesis of thienopyrimidine core to
dines with different substituents at C-2 and C-4 positions which
showed potent in vitro anticancer activity against human colon
carcinoma (HCT-116) cell line [21e23]. In the same direction to
further explore the potential of thieno[2,3-d]pyrimidines fused to
8-membered cycloalkyl ring as anticancer compounds we decided
in this work to prepare new hexahydrocycloocta [4,5]thieno[2,3-d]
pyrimidines by introducing different heteroaryl groups at C-2 po-
sition [2-pyridyl or 4-pyridyl] and different substituents at C-4
position to substantiate the effect of such substitutions on the
anticancer activity against a panel of 60 human tumor cell lines
provided by US National Cancer Institute.
2. Results and discussion
2.1. Chemistry
The synthesis of the target compounds is outlined in Scheme 1. 2-
amino-4,5,6,7,8,9-hexahydrocycloocta [4,5]thiophene-3-carboxamide
(3) our primary starting compound was prepared via two steps pro-
cedure which involved reacting cyclooctanone with cyanoacetamide 1
to afford
reacted with sulphur and diethylamine [24]. Reacting compound
with the appropriate pyridine carboxyaldehyde in dry
a-cyano-a-cyclooctylideneacetamide (2) which was then
* Corresponding author. Tel.: þ20 23639307; fax: þ20 23635140.
E-mail address: asmaa_kassab2001@yahoo.com (A.E. Kassab).
3
0223-5234/$ e see front matter Ó 2013 Elsevier Masson SAS. All rights reserved.
http://dx.doi.org/10.1016/j.ejmech.2013.02.011

A.E. Kassab, E.M. Gedawy / European Journal of Medicinal Chemistry 63 (2013) 224e230
225
F
NH
O
NH
Cl
O
O
N
O
O
O
O
N
N
N
N
Erlotinib (TarcevaTM
)
Gefitinib (IressaTM
)
O
NH
O
N
N
H3CO
N
N
O
N
Tandutinib (MLN518)
Fig. 1. Examples of 4-anilinoquinazoline compounds are potent anticancer drugs.
dimethylformamide in the presence of concentrated hydrochloric acid
resulted in 2-pyridyl hexahydrocycloocta [4,5]thieno[2,3-d]pyr-
imidin-4-(3H)-ones 4a,b. The IR spectra of 4a,b showed the presence
of an absorption band at 3097 and 3163 cmꢀ1 corresponding to NH
group. Whereas, the C]O group appeared as an absorption band at
1678 and 1658 cmꢀ1 respectively. Further evidence was obtained from
in vitro observed data, it was found that compounds 4a, 6a, 7a, 7d
and 7g showed potent anticancer activity at low concentrations
against most of the used human tumor cell lines comparable with
doxorubicin as standard potent anticancer drug (average log10 GI50
over all cell lines ¼ ꢀ6.85). Compound 4b was selective against SNB-
75 (CNS cancer) log10 GI50 ¼ ꢀ5.57. It worth mentioning that com-
pound 5a was selective against UO-3, DU-145 and T-47D cell lines
(renal, prostate and breast cancers, respectively) showing growth
inhibition percentages 83.57, 77.45 and 57.81 at a single dose test.
Interestingly, compound 7e exhibited promising selectivity against
13 cell lines representing leukemia and cancers of lung, colon, cen-
tral nervous system, ovary, kidney, prostate as well as breast
showing growth inhibition percentages between 54.05 and 89.23.
It was found that compounds (4a,b) with carbonyl group at C-4
position exhibited potent anticancer activity and 4a was more
potent than 4b. Introduction of a chloro group at C-4 position in
compounds (5a,b) resulted in a marked decrease in the activity, on
the other hand, compound 6a bearing a hydrazinyl moiety at po-
sition 4 was one of the most potent test compounds (these obser-
vations were in accordance with the previously reported work
[22]). Among the 4-substitutedaminothieno[2,3-d]pyrimidines
7aeh compound 7a with C-2 2-pyridyl and morpholinyl moiety at
C-4 position showed the most potent anticancer activity. Also
compounds 7d and 7g showed significant anticancer activity.
In the present work, we can conclude:
1H NMR spectra that showed an exchangeable singlet signals at
and 12.73 ppm corresponding to NH protons.
d 11.63
On refluxing 4a,b with phosphorus oxychloride, the corre-
sponding 4-chloro derivatives 5a,b were obtained. The IR spectra of
compounds 5a,b lacked the presence of NH and C]O absorption
bands, where as the 1H NMR spectra of these compounds revealed
the disappearance of the signal due to NH proton which confirmed
the success of chlorination.
Reacting compounds 5a,b with hydrazine hydrate in ethanol
afforded the 4-hydrazinyl derivatives 6a,b. The 1H NMR spectra of
6a and 6b showed NH2 and NH peaks as exchangeable singlet
signals at
d 4.64, 8.31 and d 4.85, 8.20 ppm respectively.
The 4-substitutedaminothieno[2,3-d]pyrimidine derivatives7aeh
wereobtainedthroughreactingcompounds5a,bwiththeappropriate
secondary amine in ethanol in the presence of catalytic amount of
triethylamine. The 1H NMR spectra of the products 7aeh revealed the
presence of expected signals corresponding to the different N-
substituted groupswhichwereindicativeforthesuccessofamination.
2.2. Anticancer activity
1 The pyridyl group at C-2 position appeared to have a consid-
erable effect on the anticancer activity since it was found that
the best activity was obtained by compounds bearing 2-pyridyl
group at the 2 position.
2 The introduction of different substituents at 4 position showed
a remarkable effect on the anticancer activity. Compounds with
carbonyl, hydrazinyl or substitutedamino group exhibited
potent anticancer activity, while those with chloro group at C-4
position were inactive.
The in vitro anticancer activity of all the newly synthesized
compounds were evaluated using a two-stage process utilizing 60
different human tumor cell lines, representing leukemia, melanoma
and cancers of lung, colon, central nervous system, ovary, kidney,
prostate as well as breast. The first stage involved the screening of all
compounds against 49 cell lines at a single dose (10ꢀ5 M). The
growth inhibition percentages obtained from the single dose test for
compounds 5a, 5b, 6b, 7b, 7c, 7e, 7f and 7h are shown in Table 1. Six
compounds 4a, 4b, 6a, 7a, 7d and 7g showed significant growth
inhibition so they were evaluated against 56 cell lines at 5 concen-
tration levels. The relationship between percentage growth and log10
of sample concentration was plotted to obtain log10 GI50 (concen-
tration required for 50% inhibition of cell growth). The log10 GI50 of
these six compounds are shown in Table 2. From the analysis of the
3 The bulkiness of substituted amino group at C-4 position
appeared to have an effect on the anticancer activity.
Further studies are required to discover the mechanism of
action and the effect of varying the substitutions at position 2 and
position 4 on optimization of the anticancer activity.

226
A.E. Kassab, E.M. Gedawy / European Journal of Medicinal Chemistry 63 (2013) 224e230
O
O
NC
+
NH2
1
O
NH2
CN
2
S/NH(C2H5)2
O
O
O
Cl
NH2
Ar
H
NH
POCl3
N
DMF/HCl
S
NH2
S
N
Ar
S
N
Ar
3
4a,b
5a,b
HN
X
NH2NH2
C2H5OH/
TEA
X
N
NH2
HN
N
N
S
S
N
Ar
N
Ar
7a-h
6a,b
Ar=
,
N
N
,
N
OCH3
X= O,
N CH3 , N
Scheme 1. The synthetic path and reagents for the preparation of the target compounds.
3. Conclusion
NMR FXQ-500 MHz spectrometers, using TMS as the internal
standard. Mass spectra were recorded on a GCMP-QP1000 EX Mass
spectrometer. Progress of the reactions were monitored by TLC
using precoated aluminum sheet silica gel MERCK 60 F254 and was
visualized by UV lamp.
Hexahydrocycloocta [4,5]thieno[2,3-d]pyrimidines represent
novel and promising class of anticancer agents. Compounds 4a, 6a,
7a, 7d and 7g showed potent anticancer activity at low concen-
trations against most of the used human tumor cell lines when
compared to doxorubicin as potent anticancer drug. The effect of
substitutions at C-2 and C-4 positions on the anticancer activity
was shown.
4.1.2. General procedure for the preparation of 2-pyridyl-
5,6,7,8,9,10-hexahydrocycloocta [4,5]thieno[2,3-d]pyrimidin-4(3H)-
ones (4a,b)
A mixture of aminoamide 3 (2.25 g, 0.01 mol) and the appropriate
pyridine carboxyaldehyde (0.03 mol) in dry dimethylformamide
(25 mL) containing concentrated hydrochloric acid (0.2 mL) was
refluxed for 24 h. The reaction mixture was cooled, filtered and the
precipitate was crystallized from the appropriate solvent.
4. Experimental
4.1. Chemistry
4.1.1. General
Melting points were obtained on a Griffin apparatus and were
uncorrected. Microanalyses for C, H and N were carried out at the
microanalytical center, Cairo University. IR spectra were recorded
4.1.2.1. 2-(2-pyridyl)-5,6,7,8,9,10-hexahydrocycloocta
[4,5]thieno
[2,3-d]pyrimidin-4(3H)-one (4a). mp 188e190 ꢁC (ethanol); yield
50%; IR (KBr) vmax: 3097 (NH), 1678 (C]O) cmꢀ1; 1H NMR (DMSO-
on a Shimadzu 435 spectrometer, using KBr discs. 1H NMR and 13
C
d6):
d 1.25e1.35 (m, 2H, CH2), 1.40e1.50 (m, 2H, CH2), 1.60e1.75 (m,
NMR spectra were performed on JOEL NMR FXQ-300 MHz and JOEL
4H, 2CH2), 2.89e2.92 (t, 2H, J ¼ 5.4 Hz, CH2), 3.09e3.11 (t, 2H,

A.E. Kassab, E.M. Gedawy / European Journal of Medicinal Chemistry 63 (2013) 224e230
227
Table 1
Growth inhibition percentages obtained from the single dose (10ꢀ5 M) test.
Panel/Cell line
Compound
5a
5b
23.77
3.12
13.63
15.59
0.89
6b
25.88
16.41
24.74
9.09
7b
7c
7e
7f
7h
Leukemia
CCRF-CEM
HL-60(TB)
K-562
MOLT-4
RPMI-8226
SR
Non-small cell lung cancer
A549/ATCC
HOP-62
27.62
3.98
20.23
32.28
4.46
2.07
5.41
4.37
30.98
e
e
e
24.65
29.28
24.54
32.16
24.06
2.53
7.55
4.82
3.42
5.99
15.19
19.98
7.6
12.38
79.22
47.89
57.54
e
23.23
16.12
28.76
12.05
15.67
33.68
21.42
e
e
e
0.18
13.37
e
ꢀ3.56
ꢀ9.05
ꢀ1.81
24.92
1.14
22.62
68.23
36.23
26.78
36.97
17.72
46.03
ꢀ0.56
ꢀ5.44
33.33
1.33
14.1
0.29
4.03
ꢀ3.13
HOP-92
e
e
e
e
NCI-H23
NCI-H322M
NCI-H460
Colon cancer
COLO 205
HCC-2998
HCT-116
HCT-15
HT29
KM12
SW-620
CNS cancer
SF-268
17.63
15.47
9.1
3.14
3.46
ꢀ2.59
7.74
3.42
ꢀ4.36
8.29
15.99
1.03
2.79
17.28
ꢀ7.78
ꢀ14.33
ꢀ2.96
8.02
ꢀ6.17
5.35
ꢀ14.58
36.56
15.54
15.77
0.67
8.45
ꢀ2.34
ꢀ16.56
ꢀ12.27
2.69
2.31
ꢀ15.99
4.13
ꢀ0.84
ꢀ8.59
23.24
20.79
17.65
19.62
ꢀ9.82
0.37
4.02
14.13
2.35
5.9
8.11
26.28
13.66
80.46
41.35
42.02
55.05
24
ꢀ1.65
ꢀ13.96
ꢀ3.45
2.1
12.8
4.95
5.76
ꢀ11.5
23.3
17.68
23.44
22.39
1.53
0.89
7.32
ꢀ1.58
13.17
ꢀ0.57
ꢀ8.49
0.47
ꢀ3.44
ꢀ1.34
ꢀ9.11
5.35
e
0.85
e
ꢀ17.76
6.22
18.8
4.06
19.48
0.97
15.83
21.94
ꢀ1.9
13.81
14.41
37.37
8.31
20.42
54.8
9.19
12.6
37.1
ꢀ1.33
6.51
2.5
11.47
ꢀ13.66
7.87
SF-295
SNB-75
SF-539
U251
15.03
2.75
2.82
9.59
6.69
ꢀ0.32
ꢀ23.78
ꢀ14.85
1.14
6.15
0.24
15.85
Melanoma
LOX IMVI
MALME-3M
M14
MDA-MB-435
SK-MEL-28
UACC-62
Ovarian cancer
IGROV1
OVCAR-3
OVCAR-4
OVCAR-5
OVCAR-8
NCI/ADR-RES
SKeOV-3
Renal cancer
786-0
32.24
5.75
13.22
8.64
10.69
4.04
5.4
7.42
5.88
2.19
11.6
ꢀ1.71
ꢀ15.42
0.03
4.93
ꢀ0.36
6.12
42.2
7.96
27.09
34.17
7.85
0
8.33
ꢀ7.69
ꢀ0.58
4.86
ꢀ6.19
ꢀ8.43
2.52
ꢀ7.52
ꢀ11.92
3.14
1.01
4.89
ꢀ4.12
6.14
4.94
10.83
ꢀ7.03
ꢀ5.08
ꢀ15.02
ꢀ5.27
15.5
9.41
41.19
5.59
36.85
10.21
ꢀ7.89
14.27
ꢀ7.94
12.86
10.65
ꢀ5.16
7.04
ꢀ5.97
14.85
ꢀ5.24
ꢀ1.87
ꢀ6.46
ꢀ7.46
ꢀ16.49
ꢀ0.84
18.72
ꢀ0.54
2.51
1.53
15.43
22.72
ꢀ1.32
15.33
18.67
0.61
6.64
ꢀ5.6
62.89
27.02
43.97
36.83
44.51
57.49
12.58
0.86
7.59
0.34
1.59
8.55
5.34
1.97
3.43
9.75
11.14
2.26
5.9
15.99
2.04
ꢀ1.5
2.44
9.44
22.71
ꢀ3.22
9.28
0.84
1.61
ꢀ18.36
120.39
165.16
e
0.46
37.24
ꢀ0.47
ꢀ4.99
e
18.15
18.55
8.69
7.27
e
32.88
33.72
17.12
15.76
e
10.04
34.81
4.94
10.69
38.39
56.07
45.88
32.24
28.9
1.77
4.79
7.05
8.66
0.87
11.09
6.97
A498
ACHN
CAKI-1
RXF 393
SN12C
UO-31
19.95
28.83
17.21
31.76
11.09
19.99
e
15.14
83.57
4.53
13.95
4.3
20.41
19.31
38.71
3.33
32.83
ꢀ12.22
55.76
16.13
Prostate cancer
PC-3
DU-145
Breast cancer
MCF7
MDA-MB-231/ATCC
HS 578T
BT-549
T-47D
MDA-MB-468
7.81
77.45
0.63
8.02
7.42
ꢀ30.41
35.43
1.4
15.29
ꢀ11.64
54.05
21.09
5.9
ꢀ4.95
44.54
2.73
45.99
ꢀ16.91
8.7
14.87
57.81
184.32
3.18
ꢀ6.99
6.64
5.87
1.46
28.53
28.1
ꢀ13.29
ꢀ20.36
19.19
50.4
6.82
6.23
10.28
24.87
5.4
89.23
22.41
6.17
2.77
57.56
30.07
2.49
2.4
ꢀ12.86
ꢀ14.13
11.7
18.15
12.06
ꢀ5.59
ꢀ17.07
26.97
0.83
ꢀ13.67
ꢀ2.76
1.1
6.79
0.16
20.15
39.65
11.82
3.79
J ¼ 5.4 Hz, CH2), 7.60e7.64 (t, 1H, J ¼ 6 Hz, ArH), 8.01e8.06 (t, 1H,
J ¼ 7.8 Hz, ArH), 8.34 (d, 1H, J ¼ 7.8 Hz, ArH), 8.74 (d, 1H, J ¼ 6 Hz,
ArH) and 11.63 (s, 1H, NH, D2O exchangeable) ppm; 13C NMR
4.1.2.2. 2-(4-pyridyl)-5,6,7,8,9,10-hexahydrocycloocta
[4,5]thieno
[2,3-d]pyrimidin-4(3H)-one (4b). mp > 300 ꢁC (n-butanol); yield
80%; IR (KBr) vmax: 3163 (NH), 1658 (C]O) cmꢀ1; 1H NMR (DMSO-
(DMSO-d6):
d
24.33, 25.24, 25.37, 26.18, 29.80, 31.45, 121.84, 126.29,
d6): d 1.20e1.30 (m, 2H, CH2), 1.35e1.45 (m, 2H, CH2), 1.55e1.65 (m,
134.16, 136.70, 137.96, 148.05, 149.06, 149.41, 157.26, 162.20,
162.75 ppm; MS [m/z, %]: 311 [Mþ, 83.66] and 78 [C5H4Nþ, 100].
Anal. Calcd for C17H17N3OS (311.39): C, 65.56; H, 5.50; N, 13.49.
Found: C, 65.59; H, 5.48; N, 13.45.
4H, 2CH2), 2.82e2.90 (m, 2H, CH2), 3.00e3.10 (m, 2H, CH2), 8.01 (d,
2H, J ¼ 5 Hz, ArH), 8.71 (d, 2H, J ¼ 5 Hz, ArH), and 12.73 (s, 1H, NH,
D2O exchangeable) ppm; 13C NMR (DMSO-d6):
d 24.93, 25.96, 26.77,
30.43, 31.94, 32.02, 115.81, 121.90, 123.12, 131.02, 148.50, 150.76,

228
A.E. Kassab, E.M. Gedawy / European Journal of Medicinal Chemistry 63 (2013) 224e230
159.08, 162.21, 168.49 ppm; MS [m/z, %]: 311 [Mþ, 100]. Anal. Calcd
for C17H17N3OS (311.39): C, 65.56; H, 5.50; N, 13.49. Found: C, 65.65;
H, 5.51; N, 13.51.
Table 2
Concentrations required for 50% inhibition of cell growth (log10 GI50).
4.1.3. General procedure for the preparation of 4-chloro-2-pyridyl-
5,6,7,8,9,10-hexahydrocycloocta [4,5]thieno[2,3-d]pyrimidines
(5a,b)
A mixture of thienopyrimidone 4a,b (0.003 mol) and phosphorus
oxychloride (15 mL) was refluxed for 1 h. The reaction mixture was
concentrated under reduced pressure then poured into ice cold
water (100 mL). The precipitated solid was filtered, washed with
water (2 ꢂ 10 mL), dried and crystallized from ethanol.
Panel/Cell line
Compound
4a 4b
6a
7a
7d
7g
Leukemia
CCRF-CEM
HL-60(TB)
K-562
MOLT-4
RPMI-8226
SR
Non-small cell lung cancer
A549/ATCC
HOP-62
ꢀ6.04 >ꢀ4.00 ꢀ5.35 ꢀ5.45 ꢀ5.09
ꢀ5.47 >ꢀ4.00 ꢀ5.35 ꢀ5.35 ꢀ4.90
ꢀ6.00 >ꢀ4.00 ꢀ5.42 ꢀ5.46 ꢀ5.45
ꢀ5.62 >ꢀ4.00 ꢀ5.44 ꢀ5.42 ꢀ5.42
ꢀ5.44 >ꢀ4.00 ꢀ5.49 ꢀ5.67 ꢀ5.08
ꢀ5.36
ꢀ5.39
ꢀ5.38
ꢀ5.49
ꢀ5.43
ꢀ5.50
ꢀ6.06 >ꢀ4.00 ꢀ5.40
e
ꢀ5.55
ꢀ5.59 >ꢀ4.00 ꢀ5.50 ꢀ5.45 ꢀ4.94
ꢀ5.20 >ꢀ4.00 ꢀ5.07 ꢀ5.02 ꢀ5.19
ꢀ5.41 >ꢀ4.00 ꢀ5.45 ꢀ6.21 ꢀ6.18
5.49
ꢀ4.94
ꢀ5.49
ꢀ5.19
ꢀ4.77
ꢀ5.26
ꢀ5.33
ꢀ4.73
4.1.3.1. 4-chloro-2-(2-pyridyl)-5,6,7,8,9,10-hexahydrocycloocta [4,5]
thieno[2,3-d]pyrimidine (5a). mp 118e120 ꢁC; yield 60%; IR (KBr)
HOP-92
vmax: 1581 (C]N), 1550 (C]C) cmꢀ1; 1H NMR (DMSO-d6):
d 1.15e
NCI-H226
NCI-H23
NCI-H322M
NCI-H460
NCI-H522
Colon Cancer
COLO 205
HCC-2998
HCT-116
HCT-15
HT29
KM12
SW-620
CNS cancer
SF-268
>ꢀ4.00 >ꢀ4.00 ꢀ4.99 ꢀ5.30 ꢀ4.98
ꢀ4.96 >ꢀ4.00 ꢀ5.05 ꢀ5.22 ꢀ4.94
ꢀ4.07 >ꢀ4.00 ꢀ5.27 ꢀ5.26 ꢀ4.88
ꢀ5.72 >ꢀ4.00 ꢀ5.38 ꢀ5.23 ꢀ4.87
>ꢀ4.00 >ꢀ4.00 ꢀ4.92 ꢀ5.23 ꢀ4.92
1.25 (m, 2H, CH2), 1.40e1.47 (m, 2H, CH2), 1.60e1.68 (m, 2H, CH2),
1.69e1.75 (m, 2H, CH2), 2.99e3.01 (t, 2H, J ¼ 5.4 Hz, CH2), 3.11e3.13
(t, 2H, J ¼ 5.4 Hz, CH2), 7.50e7.52 (t, 1H, J ¼ 3.85 Hz, ArH), 7.94e7.96
(t, 1H, J ¼ 8.35 Hz, ArH), 8.36 (d, 1H, J ¼ 8.35 Hz, ArH) and 8.73 (d,
1H, J ¼ 3.85 Hz, ArH) ppm; 13C NMR (DMSO-d6):
d 24.93, 25.48,
ꢀ5.23 >ꢀ4.00 ꢀ5.29 ꢀ5.03 ꢀ5.31
ꢀ4.58 >ꢀ4.00 ꢀ5.08 ꢀ5.16 ꢀ5.41
ꢀ5.25
ꢀ4.99
ꢀ5.49
ꢀ5.37
ꢀ5.30
ꢀ5.29
ꢀ5.29
26.20, 27.89, 30.43, 31.74, 124.11, 125.74, 127.13, 129.73, 137.74,
144.21, 150.34, 152.99, 153.45, 157.18, 169.24 ppm; MS [m/z, %]: 331
[(M þ 2)þ, 9.95], 329 [Mþ, 26.19] and 78 [C5H4Nþ, 100]. Anal. Calcd
for C17H16ClN3S (329.83): C, 61.90; H, 4.89; N, 12.73. Found: C,
62.40; H, 4.93; N, 12.84.
ꢀ5.86
e
ꢀ5.45 ꢀ5.43 ꢀ5.29
ꢀ5.63 >ꢀ4.00 ꢀ5.95 ꢀ5.47 ꢀ4.95
ꢀ5.11 >ꢀ4.00 ꢀ5.26 ꢀ5.22 ꢀ5.32
ꢀ5.39 >ꢀ4.00 ꢀ5.19 ꢀ5.37 ꢀ4.87
ꢀ5.31 >ꢀ4.00 ꢀ5.34 ꢀ5.05 ꢀ4.92
ꢀ5.06 >ꢀ4.00 ꢀ5.14 ꢀ5.05 ꢀ5.27 >ꢀ4.00
4.1.3.2. 4-chloro-2-(4-pyridyl)-5,6,7,8,9,10-hexahydrocycloocta [4,5]
thieno[2,3-d]pyrimidine (5b). mp 194e196 ꢁC; yield 65%; IR (KBr)
SF-295
SNB-19
SNB-75
SF-539
e
e
e
e
e
e
ꢀ5.47
ꢀ5.08
e
e
e
e
vmax: 1631 (C]N), 1550(C]C) cmꢀ1 1H NMR (DMSO-d6):
; d 1.15e
ꢀ4.07
ꢀ5.57 ꢀ4.90 ꢀ5.15 ꢀ5.86
ꢀ4.58
ꢀ4.17
ꢀ5.37
1.25 (m, 2H, CH2), 1.37e1.50 (m, 2H, CH2), 1.65e1.75 (m, 4H,
2CH2), 3.03e3.06 (t, 2H, J ¼ 6.1 Hz, CH2), 3.12e3.13 (t, 2H, J ¼ 6.1 Hz,
CH2), 8.63 (d, 2H, J ¼ 5 Hz, ArH) and 8.97 (d, 2H, J ¼ 5 Hz, ArH) ppm;
ꢀ5.11 >ꢀ4.00 ꢀ4.96 ꢀ4.89 ꢀ5.08
ꢀ5.67 >ꢀ4.00 ꢀ5.35 ꢀ5.36 ꢀ5.03
U251
Melanoma
LOX IMVI
MALME-3M
M14
MDA-MB-435
SK-MEL-2
SK-MEL-28
SK-MEL-5
UACC-257
UACC-62
Ovarian Cancer
IGROV1
OVCAR-3
OVCAR-4
OVCAR-5
OVCAR-8
NCI/ADR-RES
SKeOV-3
Renal cancer
786-0
A498
ACHN
CAKI-1
RXF 393
SN12C
TK-10
UO-31
Prostate cancer
PC-3
DU-145
ꢀ5.96 >ꢀ4.00 ꢀ5.21 ꢀ5.28 ꢀ5.08
ꢀ5.20
13C NMR (DMSO-d6):
d 24.40, 24.92, 25.58, 27.38, 29.89, 31.21,
e
>ꢀ4.00
e
ꢀ5.48
e
e
122.57, 127.37, 129.67, 145.16, 145.67, 147.67, 152.64, 154.14,
168.62 ppm; MS [m/z, %]: 331 [(M þ 2)þ, 60.31], 329 [Mþ,100]. Anal.
Calcd for C17H16ClN3S (329.83): C, 61.90; H, 4.89; N, 12.73. Found: C,
61.88; H, 4.88; N, 12.73.
ꢀ5.39 >ꢀ4.00 ꢀ5.26 ꢀ5.38 ꢀ5.27
ꢀ5.27
ꢀ5.26
ꢀ4.74
>ꢀ4.00 >ꢀ4.00 ꢀ5.29 ꢀ5.32 ꢀ4.93
>ꢀ4.00 >ꢀ4.00 ꢀ4.84 ꢀ5.10 ꢀ4.84
ꢀ5.22 >ꢀ4.00 ꢀ4.83 ꢀ4.96 ꢀ4.90 >ꢀ4.00
ꢀ5.53 >ꢀ4.00 ꢀ5.50 ꢀ5.70 ꢀ5.10
>ꢀ4.00 >ꢀ4.00 ꢀ5.18 ꢀ4.96 ꢀ4.97
ꢀ5.26 >ꢀ4.00 ꢀ5.23 ꢀ5.46 ꢀ4.96
ꢀ5.55
ꢀ5.14
ꢀ5.16
4.1.4. General procedure for the preparation of 4-hydrazinyl-2-
pyridyl-5,6,7,8,9,10-hexahydrocycloocta [4,5]thieno[2,3-d]
pyrimidines (6a,b)
A mixture of chloro derivative 5a,b (0.002 mol) and hydrazine
hydrate (99%, 0.62 g, 0.012 mol) in absolute ethanol (20 mL) was
refluxed for 6 h. The reaction mixture was then cooled and the
separated solid was filtered, dried and crystallized from ethanol.
ꢀ5.40 >ꢀ4.00 ꢀ5.38 ꢀ5.50 ꢀ5.09
ꢀ5.07 >ꢀ4.00 ꢀ5.27 ꢀ5.34 ꢀ4.87
ꢀ5.27
ꢀ5.06
ꢀ5.07
ꢀ4.29
ꢀ5.28
ꢀ5.32
ꢀ5.23
e
ꢀ4.98 ꢀ5.31 ꢀ4.90
4.04 >ꢀ4.00 ꢀ5.06 ꢀ4.99 ꢀ4.97
ꢀ5.36 >ꢀ4.00 ꢀ5.36 ꢀ5.29 ꢀ4.91
ꢀ6.02
e
e
ꢀ6.34 ꢀ6.08 ꢀ5.09
>ꢀ4.00
ꢀ4.51 ꢀ5.03 ꢀ4.92 >ꢀ4.00
4.1.4.1. 4-hydrazinyl-2-(2-pyridyl)-5,6,7,8,9,10-hexahydrocycloocta
ꢀ5.25 >ꢀ4.00 ꢀ5.25 ꢀ4.93 ꢀ5.32
ꢀ5.88 >ꢀ4.00 ꢀ5.73 ꢀ5.78 ꢀ5.98
ꢀ5.28 >ꢀ4.00 ꢀ5.31 ꢀ5.35 ꢀ5.10
ꢀ5.48 >ꢀ4.00 ꢀ6.24 ꢀ5.64 ꢀ5.46
ꢀ5.07
ꢀ5.48
ꢀ5.29
ꢀ5.27
[4,5]thieno[2,3-d]pyrimidine (6a). mp 172e174 ꢁC; yield 76%; IR
(KBr) vmax: 3363, 3300 (NH/NH2), 1593 (C]N) cmꢀ1 1H NMR
;
(DMSO-d6): d 1.30e1.37 (m, 2H, CH2), 1.40e1.45 (m, 2H, CH2), 1.68e
ꢀ5.13 >ꢀ4.00 ꢀ5.17 ꢀ5.08 ꢀ5.39 >ꢀ4.00
1.78 (m, 4H, 2CH2), 2.88e3.01 (m, 4H, 2CH2), 4.64 (s, 2H, NH2, D2O
exchangeable), 7.51e7.55 (m, 1H, ArH), 7.90e8.00 (t, 1H, ArH), 8.31
(s, 1H, NH, D2O exchangeable), 8.40e8.50 (m, 1H, ArH) and 8.65e
ꢀ5.20 >ꢀ4.00 ꢀ5.20 ꢀ5.30 ꢀ5.25
ꢀ5.22
>ꢀ4.00 >ꢀ4.00 ꢀ5.04 ꢀ5.03 ꢀ5.17 >ꢀ4.00
ꢀ5.23 >ꢀ4.00 ꢀ5.99 ꢀ5.93 ꢀ5.80
ꢀ5.48
8.80 (m, 1H, ArH) ppm; 13C NMR (DMSO-d6):
d 23.19, 25.18, 25.46,
ꢀ5.39 >ꢀ4.00 ꢀ5.40 ꢀ5.57 ꢀ5.33
ꢀ5.17 >ꢀ4.00 ꢀ5.02 ꢀ5.16 ꢀ4.90
ꢀ5.48
ꢀ4.55
26.48, 29.50, 30.91, 117.02, 119.20, 119.83, 125.06, 129.88, 137.19,
141.31, 150.15, 156.80, 157.11, 168.43 ppm; MS [m/z, %]: 325 [Mþ,
4.11], 57 [100]. Anal. Calcd for C17H19N5S (325.42): C, 62.73; H, 5.88;
N, 21.52. Found: C, 62.63; H, 5.87; N, 21.48.
Breast cancer
MCF7
MDA-MB-231/ATCC
HS 578T
BT-549
5.61 >ꢀ4.00 ꢀ5.42 ꢀ5.52 ꢀ5.18
ꢀ5.06 >ꢀ4.00 ꢀ4.90 ꢀ5.13 ꢀ5.37
ꢀ5.19 >ꢀ4.00 ꢀ4.96 ꢀ5.47 ꢀ5.64
ꢀ5.25 >ꢀ4.00 ꢀ5.01 ꢀ5.36 ꢀ5.20
ꢀ5.50
ꢀ4.30
ꢀ4.24
ꢀ5.16
ꢀ5.20
ꢀ5.39
4.1.4.2. 4-hydrazinyl-2-(4-pyridyl)-5,6,7,8,9,10-hexahydrocycloocta
T-47D
MDA-MB-468
ꢀ5.26
e
ꢀ5.30 ꢀ5.35 ꢀ4.87
[4,5]thieno[2,3-d]pyrimidine (6b). mp 212e214 ꢁC; yield 80%; IR
ꢀ5.78 >ꢀ4.00 ꢀ5.44 ꢀ5.38 ꢀ5.28
(KBr) vmax: 3305, 3300 (NH/NH2), 1593 (C]N) cmꢀ1 1H NMR
;
(DMSO-d6):
d 1.10e1.15 (m, 2H, CH2), 1.37e1.45 (m, 2H, CH2), 1.65e
1.75 (m, 4H, 2CH2), 2.85e2.90 (t, 2H, J ¼ 6 Hz, CH2), 3.03e3.07 (t,

A.E. Kassab, E.M. Gedawy / European Journal of Medicinal Chemistry 63 (2013) 224e230
229
2H, J ¼ 6 Hz, CH2), 4.85 (s, 2H, NH2, D2O exchangeable), 8.20 (s, 1H,
NH, D2O exchangeable), 8.31 (d, 2H, J ¼ 5 Hz, ArH) and 8.67 (d, 2H,
N(CH3)eCH2e), 3.70e3.90 (m, 4H, eCH2eNeCH2e), 8.27 (d, 2H,
J ¼ 6 Hz, ArH) and 8.74 (d, 2H, J ¼ 6 Hz, ArH) ppm; 13C NMR (DMSO-
J ¼ 5 Hz, ArH) ppm; 13C NMR (DMSO-d6):
d
24.32, 24.88, 25.57,
d6): d 24.55, 24.94, 25.47, 26.99, 30.79, 31.21, 42.29, 47.46, 51.90,
27.06, 29.99, 31.55, 114.22, 121.62, 129.36, 136.51, 144.99, 149.98,
155.29, 157.87, 164.49 ppm; MS [m/z, %]: 325 [Mþ, 0.94] and 295
[C17 H17N3Sþ, 100]. Anal. Calcd for C17H19N5S (325.42): C, 62.73; H,
5.88; N, 21.52. Found: C, 62.47; H, 5.85; N, 21.43.
119.14, 121.45, 129.66, 140.44, 144.27, 150.28, 154.31, 161.06,
169.02 ppm; MS [m/z, %]: 393 [Mþ, 6.08] and 70 [C4H8Nþ, 100].
Anal. Calcd for C22H27N5S (393.54): C, 67.13; H, 6.91; N, 17.79.
Found: C, 67.03; H, 6.90; N, 17.76.
4.1.5. General procedure for the preparation of 2-pyridyl-4-
substitutedamino-5,6,7,8,9,10-hexahydrocycloocta [4,5]thieno[2,3-
d] pyrimidines (7aeh)
A mixture of 5a,b (0.001 mol), the selected secondary amine
(0.001 mol) and triethylamine (0.36 mL, 0.003 mol) in absolute
ethanol (12 mL) was heated under reflux for 15 h. The separated
solid after cooling was filtered, dried and crystallized from ethanol.
4.1.5.5. 4-(4-phenylpiperazin-1-yl)-2-(2-pyridyl)-5,6,7,8,9,10-
hexahydrocycloocta [4,5]thieno[2,3-d]pyrimidine (7e). mp 128e
130 ꢁC; yield 80%; IR (KBr) vmax: 1597 (C]N), 1550 (C]C) cmꢀ1
;
1H NMR (DMSO-d6):
d 1.05e1.15 (m, 2H, CH2), 1.35e1.45 (m, 2H,
CH2), 1.53e1.60 (m, 2H, CH2), 1.65e1.75 (m, 2H, CH2), 2.93e3.00 (m,
2H, CH2), 3.05e3.10 (m, 2H, CH2), 3.15e3.25 (t, 4H, J ¼ 5.4 Hz, e
CH2eN(C6H5)eCH2e), 3.50e3.60 (m, 4H, eCH2eNeCH2e), 6.79e
6.83 (t, 1H, J ¼ 7.2 Hz, ArH), 7.00 (d, 2H, J ¼ 8.4 Hz, ArH), 7.21e
7.26 (t, 2H, J ¼ 7.2 Hz, ArH), 7.46e7.50 (t, 1H, J ¼ 6 Hz, ArH),
7.92e7.97 (t, 1H, J ¼ 7.8 Hz, ArH), 8.43 (d, 1H, J ¼ 7.8 Hz, ArH) and
4.1.5.1. 4-(morpholin-4-yl)-2-(2-pyridyl)-5,6,7,8,9,10-
hexahydrocycloocta [4,5]thieno[2,3-d]pyrimidine (7a). mp 120e
122 ꢁC; yield 79%; IR (KBr) vmax: 1581 (C]N), 1550 (C]C) cmꢀ1
;
8.73 (d, 1H, J ¼ 6 Hz, ArH) ppm; 13C NMR (DMSO-d6):
d 24.47, 24.92,
1H NMR (DMSO-d6):
d
1.05e1.10 (m, 2H, CH2), 1.35e1.40 (m, 2H,
25.50, 26.98, 30.66, 31.21, 42.40, 45.32, 47.99, 50.34, 115.59, 115.90,
119.16, 119.80, 120.51, 123.34, 124.42, 128.86, 128.97, 129.57, 136.78,
139.55, 149.94, 150.76, 156.07, 162.17, 168.15 ppm; MS [m/z, %]: 455
[Mþ, 8.05], 323 [C18H19N4Sþ, 100]. Anal. Calcd for C27H29N5S
(455.60): C, 71.17; H, 6.41; N, 15.37. Found: C, 71.24; H, 6.42; N,
15.38.
CH2), 1.50e1.60 (m, 2H, CH2), 1.65e1.70 (m, 2H, CH2), 2.90e2.95 (m,
2H, CH2), 3.00e3.05 (m, 2H, CH2), 3.35e3.45 (m, 4H, eCH2eNe
CH2e), 3.70e3.80 (m, 4H, eCH2eOeCH2e), 7.42e7.50 (t, 1H,
J ¼ 6 Hz, ArH), 7.90e7.92 (t, 1H, J ¼ 7.65 Hz, ArH), 8.38 (d, 1H,
J ¼ 7.65 Hz, ArH) and 8.70 (d, 1H, J ¼ 6 Hz, ArH) ppm; 13C NMR
(DMSO-d6):
d 24.50, 24.94, 25.51, 26.99, 30.72, 31.24, 50.92, 65.77,
118.95, 123.39, 124.46, 129.56, 136.80, 139.65, 149.44, 154.66, 156.15,
162.18, 168.20; MS [m/z, %]: 380 [Mþ, 59.33], 337 [100]. Anal. Calcd
for C21H24N4OS (380.49): C, 66.28; H, 6.35; N, 14.72. Found: C,
66.04; H, 6.33; N, 14.67.
4.1.5.6. 4-(4-phenylpiperazin-1-yl)-2-(4-pyridyl)-5,6,7,8,9,10-
hexahydrocycloocta [4,5]thieno[2,3-d]pyrimidine (7f). mp 198e
200 ꢁC; yield 91%; IR (KBr) vmax: 1597 (C]N), 1543 (C]C) cmꢀ1
;
1H NMR (DMSO-d6):
d 1.05e1.10 (m, 2H, CH2), 1.35e1.45 (m, 2H,
CH2), 1.55e1.60 (m, 2H, CH2), 1.65e1.70 (m, 2H, CH2), 2.95e3.00 (m,
2H, CH2), 3.01e3.05 (m, 2H, CH2), 3.15e3.17 (t, 4H, J ¼ 5.4 Hz, e
CH2eN(C6H5)eCH2e), 3.54e3.60 (m, 4H, eCH2eNeCH2e), 6.80e
6.83 (t, 1H, J ¼ 8.4 Hz, ArH), 6.94 (d, 2H, J ¼ 8.4 Hz, ArH), 7.20e
7.23 (t, 2H, J ¼ 8.4 Hz, ArH), 8.23 (d, 2H, J ¼ 6.1 Hz, ArH) and 8.70
4.1.5.2. 4-(morpholin-4-yl)-2-(4-pyridyl)-5,6,7,8,9,10-
hexahydrocycloocta [4,5]thieno[2,3-d]pyrimidine (7b). mp 145e
147 ꢁC; yield 67%; IR (KBr) vmax: 1597 (C]N), 1546 (C]C) cmꢀ1
;
1H NMR (DMSO-d6):
d 1.05e1.15 (m, 2H, CH2), 1.35e1.45 (m, 2H,
CH2), 1.50e1.60 (m, 2H, CH2), 1.65e1.70 (m, 2H, CH2), 2.90e3.05 (m,
4H, 2CH2), 3.35e3.45 (m, 4H, eCH2eNeCH2e), 3.75e3.85 (m, 4H, e
CH2eOeCH2e), 8.23 (d, 2H, J ¼ 6 Hz, ArH) and 8.71 (d, 2H, J ¼ 6 Hz,
(d, 2H, J ¼ 6.1 Hz, ArH) ppm; 13C NMR (DMSO-d6):
d 24.50, 24.93,
25.30, 27.03, 30.70, 31.25, 42.48, 45.38, 48.02, 50.29, 115.69, 115.95,
119.25, 119.87, 121.42, 128.93, 129.02, 129.84, 139.99, 149.98, 150.25,
154.42, 162.20 ppm; MS [m/z, %]: 455 [Mþ, 9.75], 69 [100]. Anal.
Calcd for C27H29N5S (455.60): C, 71.17; H, 6.41; N, 15.37. Found: C,
71.10; H, 6.40; N, 15.35.
ArH) ppm; 13C NMR (DMSO-d6):
d 24.51, 24.91, 25.51, 27.01, 30.72,
31.25, 50.83, 65.74, 119.16, 121.41, 129.74, 140.05, 144.41, 150.25,
154.42, 162.09, 167.98 ppm; MS [m/z, %]: 380 [Mþ, 100]. Anal. Calcd
for C21H24N4OS (380.49): C, 66.28; H, 6.35; N, 14.72. Found: C,
66.22; H, 6.35; N, 14.71.
4.1.5.7. 4-[4-(4-methoxyphenyl)-piperazin-1-yl]-2-(2-pyridyl)-5,6,7,
8,9,10-hexahydrocycloocta [4,5]thieno[2,3-d]pyrimidine (7g). mp 158e
160 ꢁC; yield 93%; IR (KBr) vmax: 1590 (C]N), 1550 (C]C) cmꢀ1; 1H
4.1.5.3. 4-(4-methylpiperazin-1-yl)-2-(2-pyridyl)-5,6,7,8,9,10-
hexahydrocycloocta [4,5]thieno[2,3-d]pyrimidine (7c). mp 118e
NMR (DMSO-d6): d 1.05e1.15 (m, 2H, CH2), 1.35e1.45 (m, 2H, CH2),
120 ꢁC; yield 64%; IR (KBr) vmax: 1630 (C]N), 1550 (C]C) cmꢀ1
;
1.50e1.60 (m, 2H, CH2), 1.62e1.70 (m, 2H, CH2), 2.90e3.00 (m, 2H,
CH2), 3.02e3.10 (m, 2H, CH2), 3.15e3.25 (m, 4H, eCH2eN(4-
CH3OC6H4)eCH2e), 3.45e3.55 (m, 4H, eCH2eNeCH2e), 3.66 (s,
3H, CH3O), 6.81 (d, 2H, J ¼ 8.4 Hz, ArH), 6.93 (d, 2H, J ¼ 8.4 Hz, ArH),
7.44e7.46 (t, 1H, J ¼ 4.6 Hz, ArH), 7.90e7.93 (t, 1H, J ¼ 7.65 Hz, ArH),
1H NMR (DMSO-d6):
d 1.05e1.15 (m, 2H, CH2), 1.35e1.45 (m, 2H,
CH2), 1.50e1.60 (m, 2H, CH2), 1.65e1.75 (m, 2H, CH2), 2.63 (s, 3H,
NCH3), 2.90e2.95 (m, 2H, CH2), 3.00e3.10 (m, 6H, CH2 and eCH2e
N(CH3)eCH2e), 3.30e3.50 (m, 4H, eCH2eNeCH2e), 7.46e7.50 (t,
1H, J ¼ 4.5 Hz, ArH), 7.92e7.97 (t, 1H, J ¼ 7.8 Hz, ArH), 8.40 (d, 1H,
J ¼ 7.8 Hz, ArH) and 8.71 (d, 1H, J ¼ 3.9 Hz, ArH) ppm; 13C NMR
8.39 (d,1H, J ¼ 7.65 Hz, ArH) and 8.69 (d,1H, J ¼ 4.6 Hz, ArH) ppm; 13
C
NMR (DMSO-d6):
d 24.51, 24.95, 25.52, 27.00, 30.70, 31.24, 42.63,
(DMSO-d6):
d
24.42, 24.93, 25.53, 26.99, 30.73, 31.23, 42.69, 49.83,
45.36, 50.37, 114.33, 115.64, 115.94, 117.64, 118.03, 119.20, 119.86,
124.47, 128.92,129.02,129.64, 139.61, 149.99,150.82,162.24 ppm; MS
[m/z, %]: 485 [Mþ, 12.53] and 323 [C18H19N4Sþ, 100]. Anal. Calcd for
53.87, 118.82, 123.36, 124.41, 129.62, 136.79, 139.44, 149.42, 154.69,
156.06, 162.05, 168.13 ppm; MS [m/z, %]: 393 [Mþ, 5.83], 323
[C18H19N4Sþ, 100]. Anal. Calcd for C22H27N5S (393.54): C, 67.13; H,
6.91; N, 17.79. Found: C, 66.98; H, 6.90; N, 17.75.
C28H31N5OS (485.63): C, 69.24; H, 6.43; N, 14.42. Found: C, 69.09; H,
6.42; N, 14.39.
4.1.5.4. 4-(4-methylpiperazin-1-yl)-2-(4-pyridyl)-5,6,7,8,9,10-
4.1.5.8. 4-[4-(4-methoxyphenyl)-piperazin-1-yl]-2-(4-pyridyl)-5,6,7,
8,9,10-hexahydrocycloocta [4,5]thieno[2,3-d]pyrimidine (7h). mp
162e164 ꢁC; yield 81%; IR (KBr) vmax: 1597 (C]N), 1550 (C]C)
hexahydrocycloocta [4,5]thieno[2,3-d]pyrimidine (7d). mp 264e
266 ꢁC; yield 68%; IR (KBr) vmax: 1597 (C]N), 1550 (C]C) cmꢀ1
;
1H NMR (DMSO-d6):
d
1.10e1.15 (m, 2H, CH2), 1.40e1.50 (m, 2H,
cmꢀ1; 1H NMR (DMSO-d6):
d 1.05e1.15 (m, 2H, CH2), 1.35e1.45 (m,
CH2), 1.55e1.65 (m, 2H, CH2), 1.70e1.75 (m, 2H, CH2), 2.77 (s, 3H,
NCH3), 2.95e3.05 (m, 2H, CH2), 3.20e3.40 (m, 6H, CH2 and eCH2e
2H, CH2), 1.55e1.60 (m, 2H, CH2), 1.65e1.75 (m, 2H, CH2), 2.95e3.04
(m, 2H, CH2), 3.05e3.10 (m, 2H, CH2), 3.20e3.30 (m, 4H, eCH2eN(4-

230
A.E. Kassab, E.M. Gedawy / European Journal of Medicinal Chemistry 63 (2013) 224e230
CH3OC6H4)eCH2e), 3.55e3.60 (m, 4H, eCH2eNeCH2e), 3.68 (s, 3H,
CH3O), 6.84 (d, 2H, J ¼ 9.3 Hz, ArH), 6.84 (d, 2H, J ¼ 9.3 Hz, ArH), 8.28
(d, 2H, J ¼ 6 Hz, ArH) and 8.74 (d, 2H, J ¼ 6 Hz, ArH) ppm; 13C NMR
Acknowledgment
We are grateful to all members of National Institute of Health,
Maryland, USA for carrying out the anticancer screening.
(DMSO-d6):
d 24.50, 24.93, 25.52, 27.04, 30.71, 31.25, 49.42, 50.41,
55.16, 114.29, 117.70, 119.23, 121.42, 129.84, 139.94, 144.45, 145.13,
150.26, 153.20, 154.41, 162.16, 167.96 ppm; MS [m/z, %]: 485 [Mþ,
12.46], 323 [C18H19N4Sþ, 100]. Anal. Calcd for C28H31N5OS (485.63):
C, 69.24; H, 6.43; N, 14.42. Found: C, 69.18; H, 6.42; N, 14.40.
Supplementary data
Supplementary data associated with this article can be found in
the online version, at http://dx.doi.org/10.1016/j.ejmech.2013.02.
011. These data include MOL files and InChiKeys of the most
important compounds described in this article.
4.2. Measurement of anticancer activity
Anticancer activity screening of the newly synthesized com-
pounds was measured in vitro utilizing 60 different human tumor
cell lines provided by US National Cancer Institute according to
previously reported standard procedure [25e27] as follows:
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