الفهرس 
CancerOnly 14 pages are availabe for public view
Abstract
According to statistics from the American Cancer Society (ACS), Cancer is the third
most lethal disease after cardiovascular diseases, infectious and parasitic diseases. It
may affect humans of all ages, regions and socioeconomic levels.
Continuous research worldwide is focusing on developing better therapeutics as
well as finding novel druggable targets for better efficacy. Small molecule tyrosine
kinase inhibitors provide attractive therapeutic targets, as they are able to block cell
signaling associated with many types of cancer.
In this study, thienopyrimidine derivatives have been designed and synthesized as
targeted EGFR/HER-2 inhibitors. The design focused on exploration of the previous
revealed SAR studies, bioisosteric modifications of the lead compounds both in
market and in clinical studies, and identification of the key interactions with the
binding site in silico.
Synthesis of the designed compounds was then accomplished & their structures
were confirmed by various spectral and microanalytical data.
This study involved the synthesis of the following reported intermediates:
1) 1-(Benzyloxy)-4-nitrobenzene (Ia)
2) 1-Fluoro-3-((4-nitrophenoxy) methyl)benzene (Ib)
3) 1-Fluoro-4-((4-nitrophenoxy) methyl)benzene (Ic)
4) 1-Chloro-4-((4-nitrophenoxy) methyl)benzene (Id)
5) 2-Chloro-1-((3-fluorobenzyl) oxy)-4-nitrobenzene (If)
6) 4-(Benzyloxy) aniline (IIa)
7) 4-((4-Chlorobenzyl) oxy) aniline (IId)
8) 4-(Benzyloxy)-3-chloroaniline (IIe)
9) 3-Chloro-4-((3-fluorobenzyl) oxy) aniline (IIf)
10) 3-Chloro-4-((4-fluorobenzyl) oxy) aniline (IIg)
11)3-Chloro-4-((4-chlorobenzyl) oxy) aniline (IIh)
12)Ethyl -5-amino-4-cyano-3-methylthiophene-2-carboxylate (III)
13) 2-Amino-4,5,6,7-tetrahydrobenzo[b]thiophene-3-carbonitrile (VI)
14)Ethyl 2-amino-4,5,6,7-tetrahydrobenzo[b]thiophene-3-carboxylate (IX)
15) 5,6,7,8-Tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidin-4(3H)-one (X)
16)4-Chloro-5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidine (XI)
17)1-(4-((5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidin-4-
yl)amino)phenyl)ethan-1-one (XII) Also, it comprised the following new intermediates:
1) 1-(Benzyloxy)-2-chloro-4-nitrobenzene (Ie)
2) 2-Chloro-1-((4-fluorobenzyl) oxy)-4-nitrobenzene (Ig)
3) 2-Chloro-1-((4-chlorobenzyl) oxy)-4-nitrobenzene (Ih)
4) 4-((3-Fluorobenzyl) oxy) aniline (IIb)
5) 4-((4-Fluorobenzyl) oxy) aniline (IIc)
6) Ethyl.(E)-4-cyano-5-(((dimethylamino)methylene)amino)-3-methylthiophene-
2-carboxylate (IV)
7) (E)-N’-(3-Cyano-4,5,6,7-tetrahydrobenzo[b]thiophen-2-yl)-N,Ndimethylformimidamide (VII)
Also, the study involved the synthesis and the characterization of the following newtargeted compounds:
1) Ethyl4-((4-(benzyloxy) phenyl) amino)-5-methylthieno [2,3-d]pyrimidine-6-
carboxylate (Va)
2) Ethyl4-((4-((3-fluorobenzyl)oxy)phenyl)amino)-5-methylthieno[2,3-
d]pyrimidine-6-carboxylate (Vb)
3) Ethyl.4-((4-((4-fluorobenzyloxy)phenyl)amino)-5-methylthieno[2,3-
d]pyrimidine-6-carboxylate (Vc)
4) Ethyl.4-((4-((4-chlorobenzyloxy)phenyl)amino)-5-methylthieno[2,3-
d]pyrimidine-6-carboxylate (Vd)
5) Ethyl4-((4-(benzyloxy)-3-chlorophenyl)amino)-5-methylthieno[2,3-
d]pyrimidine-6-carboxylate (Ve)
6) Ethyl.4-((3-fluorobenzyloxy)-3-chlorophenyl)amino)-5-methylthieno[2,3-
d]pyrimidine-6-carboxylate (Vf)
7) Ethyl.4-((4-fluorobenzyloxy)-3-chlorophenyl)amino)-5-methylthieno[2,3-
d]pyrimidine-6-carboxylate (Vg)
8) Ethyl.4-((4-chlorobenzyloxy)-3-chlorophenyl)amino)-5-methylthieno[2,3-
d]pyrimidine-6-carboxylate (Vh)
9) N-(4-(Benzyloxy)phenyl)-5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-d]
pyrimidin-4-amine(VIIIa)
10) N-(4-(3-Fluorobenzyloxy)phenyl)-5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-
d]pyrimidin-4-amine (VIIIb) 11) N-(4-(4-Fluorobenzyloxy)phenyl)-5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-
d]pyrimidin-4-amine (VIIIc)
12) N-(4-(4-Chlorobenzyloxy)phenyl)-5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-
d]pyrimidin-4-amine (VIIId)
13) N-(4-(Benzyloxy)-3-chlorophenyl)-5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-
d]pyrimidin-4-amine (VIIIe)
14) N-(3-Chloro-4-((3-fluorobenzyl)oxy)phenyl)-5,6,7,8-tetrahydrobenzo[4,5]
thieno[2,3-d]pyrimidin-4-amine (VIIIf)
15) N-(3-Chloro-4-((4-fluorobenzyl)oxy)phenyl)-5,6,7,8-tetrahydrobenzo[4,5]
thieno[2,3-d]pyrimidin-4-amine (VIIIg)
16) N-(3-Chloro-4-((4-chlorobenzyl)oxy)phenyl)-5,6,7,8-tetrahydrobenzo[4,5]
thieno [2,3-d]pyrimidin-4-amine (VIIIh)
17) (E)-3-(4-Fluorophenyl)-1-(4-((5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidin-
4-yl)amino)phenyl)prop-2-en-1-one (XIIIa)
18) (E)-3-(4-Chlorophenyl)-1-(4-((5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-
d]pyrimidin-4-yl)amino)phenyl)prop-2-en-1-one (XIIIb)
19) (E)-3-(4-Bromophenyl)-1-(4-((5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-
d]pyrimidin-4-yl)amino)phenyl)prop-2-en-1-one (XIIIc)
20) (E)-3-(4-Methoxyphenyl)-1-(4-((5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-
d]pyrimidin-4-yl)amino)phenyl)prop-2-en-1-one (XIIId)
21) (E)-3-(3,4-Dimethoxyphenyl)-1-(4((5,6,7,8−tetrahydrobenzo[4,5]thieno[2,3-
d]pyrimidin-4-yl)amino)phenyl)prop-2-en-1-one (XIIIe)
22) 5-(4-fluorophenyl)-3-(4-((5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidin-
4-yl)amino)phenyl)-4,5-dihydro-1H-pyrazole-1-carbothioamide (XIVa)
23) 5-(4-chlorophenyl)-3-(4-((5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidin-
4-yl)amino)phenyl)-4,5-dihydro-1H-pyrazole-1-carbothioamide (XIVb)
24) 5-(4-bromophenyl)-3-(4-((5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidin-
4-yl)amino)phenyl)-4,5-dihydro-1H-pyrazole-1-carbothioamide (XIVc)
25) 5-(4-methoxyphenyl)-3-(4-((5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-
d]pyrimidin-4-yl)amino)phenyl)-4,5-dihydro-1H-pyrazole-1-carbothioamide
(XIVd) 26) 5-(3,4-dimethoxyphenyl)-3-(4-((5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-
d]pyrimidin-4-yl)amino)phenyl)-4,5-dihydro-1H-pyrazole-1-carbothioamide
(XIVe)
27) 4-(4-Fluorophenyl)-2-imino-6-(4-((5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-
d]pyrimidin-4-yl)amino)phenyl)-1,2-dihydropyridine-3-carbonitrile (XVa)
28) 4-(4-Chlorophenyl)-2-imino-6-(4-((5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-
d]pyrimidin-4-yl)amino)phenyl)-1,2-dihydropyridine-3-carbonitrile (XVb)
29) 4-(4-Bromophenyl)-2-imino-6-(4-((5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-
d]pyrimidin-4-yl)amino)phenyl)-1,2-dihydropyridine-3-carbonitrile (XVc)
30) 2-Imino-4-(4-methoxyphenyl)-6-(4((5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-
d]pyrimidin-4-yl)amino)phenyl)-1,2-dihydropyridine-3-carbonitrile (XVd)
31) 4-(4-Fluorophenyl)-2-oxo-6-(4-((5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-
d]pyrimidin-4-yl)amino)phenyl)-1,2-dihydropyridine-3-carbonitrile (XVIa)
32) 4-(4-Chlorophenyl)-2-oxo-6-(4-((5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-
d]pyrimidin-4-yl)amino)phenyl)-1,2-dihydropyridine-3-carbonitrile (XVIb)
33) 4-(4-Bromophenyl)-2-oxo-6-(4-((5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-
d]pyrimidin-4-yl)amino)phenyl)-1,2-dihydropyridine-3-carbonitrile (XVIc)
34) 4-(4-Methoxyphenyl)-2-oxo-6-(4-((5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-
d]pyrimidin-4-yl)amino)phenyl)-1,2-dihydropyridine-3-carbonitrile (XVId)
Preliminary molecular docking was performed using C-DOCKER protocol in
Discovery Studio 4.0 Software, it was attempted to predict the binding mode of the
targeted compounds. The biological evaluation was accomplished through testing both
antiproliferative activity and enzyme inhibition activity. The enzymatic assay was
performed in BPS Bioscience Corporation; USA. The enzymatic activity of the
synthesized compounds was evaluated against EGFR/HER-2 tyrosine kinase at
10 μM concentration. Most of the synthesized compounds showed good to potent
EGFR/HER-2 inhibitory activity in particular Va, Vf, VIIIa, VIIIb, VIIIe, VIIIf and
VIIIg. The tetrahydrobenzothienopyrimidine derivative bearing a 3-chloro-4-(3-
fluorobenzyloxy) aniline (VIIIf) exhibited 100% inhibition on both kinases.
Furthermore, five compounds (VIIIa, VIIIb, VIIIe, VIIIf, VIIIg) demonstrated highly
potent dose-related EGFR/HER-2 inhibition with IC50 values in submicromolar range
which exhibited IC50 values of 1.2, 0.6, 0.3, 0.2, 0.4 µM and 8.2, 3.4, 1.3, 0.5, 2.7 µM on
EGFR and HER-2 respectively.
Twelve of the final Compounds (Vb, Vc , Vd, Vf, Vh, VIIIa, VIIIb, VIIIc, VIIIe, VIIIf,
VIIIg, VIIIh) were selected by the National Cancer Institute “NCI” for single dose
screening program at 10 μM in the full NCI 60 cell panel. The thieno[2,3-
d]pyrimidine-based derivative (VIIIf) showed remarkably the lowest cell growth
promotion, hence good anti-proliferative activity against specific cell lines.
Finally, Molecular docking was performed using C-DOCKER protocol in Discovery
Studio 4.0 Software, it was attempted to investigate the binding mode of the targeted
compounds and interpret their variable inhibitory activity. Computer aided ADMET
study was also performed using Discovery Studio 2.5 Software.