Computer-aided design, synthesis, and biological evaluation of new indole-2-carboxamide derivatives as PI3Kα/EGFR inhibitors

Article abstract of DOI:10.1016/j.bmcl.2016.04.011

Structure-based drug design and molecular modeling were employed to identify a new series of indole-2-carboxamides as potential anticancer agents. These compounds were synthesized and characterized with the aid of several spectroscopic techniques, such as

Full text of DOI:10.1016/j.bmcl.2016.04.011

Accepted Manuscript
Computer-aided design, synthesis, and biological evaluation of new indole-2-
carboxamide derivatives as PI3Kα/EGFR inhibitors
Kamal Sweidan, Dima A. Sabbah, Sanaa Bardaweel, Khadeja Abu Dush,
Ghassan Abu Sheikha, Mohammad S. Mubarak
PII:
S0960-894X(16)30372-9
DOI:
http://dx.doi.org/10.1016/j.bmcl.2016.04.011
BMCL 23773
Reference:
To appear in:
Bioorganic & Medicinal Chemistry Letters
Received Date:
Revised Date:
Accepted Date:
16 January 2016
3 April 2016
5 April 2016
Please cite this article as: Sweidan, K., Sabbah, D.A., Bardaweel, S., Dush, K.A., Sheikha, G.A., Mubarak, M.S.,
Computer-aided design, synthesis, and biological evaluation of new indole-2-carboxamide derivatives as PI3Kα/
EGFR inhibitors, Bioorganic & Medicinal Chemistry Letters (2016), doi: http://dx.doi.org/10.1016/j.bmcl.
2
016.04.011
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Computer-Aided Design, Synthesis, and Biological Evaluation of New Indole-2-Carboxamide
Derivatives as PI3Kα/EGFR Inhibitors
a,
b,
c
a
c
Kamal Sweidan *, Dima A. Sabbah *, Sanaa Bardaweel , Khadeja Abu Dush , Ghassan Abu Sheikha ,
a
Mohammad S. Mubarak

Bioorganic & Medicinal Chemistry Letters
Computer-aided design, synthesis, and biological evaluation of new indole-2-
carboxamide derivatives as PI3Kα/EGFR inhibitors
a,
b,
c
a
c
Kamal Sweidan *, Dima A. Sabbah *, Sanaa Bardaweel , Khadeja Abu Dush , Ghassan Abu Sheikha ,
a
Mohammad S. Mubarak
a
Department of Chemistry, The University of Jordan, Amman 11942, Jordan
b
Faculty of Pharmacy, Al-Zaytoonah University of Jordan, Amman, 11733, Jordan
c
Department of Pharmaceutical Sciences, Faculty of Pharmacy, The University of Jordan, Amman 11942, Jordan
ARTICLE INFO
ABSTRACT
Article history:
Received
Revised
Accepted
Available online
Structure-based drug design and molecular modeling were employed to identify a new series of
indole-2-carboxamides as potential anticancer agents. These compounds were synthesized and
characterized with the aid of several spectroscopic techniques, such as FT-IR, NMR, and mass
spectrometry as well as by elemental analysis. Molecular docking studies confirmed that the
newly synthesized compounds accommodate PI3Kα and EGFR kinase catalytic sites and form
H-bonding with the key binding residues. The antitumor activity of these new compounds
against an array of cancer cell lines (human colon carcinoma (HCT116), leukemia (K562), and
breast cancer (MDA231) was evaluated. Results revealed that these compounds were selective
against the kinase domain, and none of them showed any inhibitory activity against K562. In
addition, results showed that compound 13 exhibited high potency in HCT116 and MDA231
with IC50 values of 19 and 15 µM, respectively. Our findings recommend that further
optimization of this series would be beneficial for colon and breast cancer treatment.
2009 Elsevier Ltd. All rights reserved.
Keywords:
PI3Kα
EGFR
HCT116
MDA231
Docking
The chemistry and medicinal study of heterocyclic compounds
have been considered a powerful approach to treat a wide range
of diseases. Indole (benzopyrrole) system has evolved as an
important heterocyclic structure due to its presence in a large
number of bioactive compounds; this nucleus was reported as the
In continuation of our ongoing search of new heterocyclic
derivatives targeting PI3Kα inhibition, we employed an adopted
16
pharmacophore model of PI3Kα inhibitors to tailor the indole
core nucleus with functionalities that match the finger print of
active PI3Kα inhibitors (Fig. 2). Moreover, we assigned the
crystal structures of PI3Kα and EGFR as target enzymes having
kinase domain to explore the structural basis of binding of this
series and identify structural features that are required to induce
an inhibitory activity.
1-3
4
7
basic system, as shown in Fig. 1, as antiviral , antidepressant
5
6,
(
1), antihyperlipidemic (2), and anti-inflammatory agents.
Furthermore, other indole derivatives (3, 4) were reported as
potent antitumor agents and were extensively studied in treating
8-13
cancer
which still remains a major threat to the human life.
Phosphatidylinositol 3-kinase (PI3Ks) enzymes are lipid kinases
that phosphorylate the hydroxyl group at the 3 position of the
inositol ring yielding phosphatidylinositol 3,4,5 triphosphates
(PIP ). PIP regulates many cellular processes such as cell
3 3
proliferation, survival, motility, and apoptosis. On the other hand,
Similarly, the carboxamide functional group occurs in many
drugs; a property that stems from its ability to serve as a
hydrogen-bond acceptor through its carbonyl group and as
14, 15
hydrogen-bond donor via its primary/secondary amides.
For the past few years, our research group has been interested
PI3Ks are negatively regulated by phosophoinositide
in the design and synthesis of small molecular weight inhibitors
phosphatases such as phosphatase and tensin homolog (PTEN)
14-20
24, 25
that target phosphatidylinositol 3-kinase (PI3Kα).
We
which dephosphorylate the 3 OH position of PIP
3
.
PI3Ks are
synthesized two novel scaffolds: N-phenyl-4-hydroxy-2-
divided into three classes (I, II, and III) based on their substrate
interaction and coding sequence. Class IA PI3K has PI3Kα
isoform encoded by PIK3CA gene. PI3Kα is amplified,
overexpressed, and mutated in a variety of human tumors
including breast, p2r6ostate, cervix, gastric, colon, and
endometrium tumors. Cancer derived "hotspots" somatic
mutations were observed in PI3Kα coding sequence and are
clustered in helical (E542K and E545K) and kinase (H1047R)
17
quinolone-3-carboxamides
as selective (H1047R) PI3Kα
19
inhibitors and derivatives of benzoin as PI3Kα inhibitors. In
addition, we have recently described the synthesis and
cytotoxicity evaluation of new benzofuran and benzothiophene
21
carboxamide models, and employed molecular docking studies
for novel heterocyclic carboxamides as potential PI3Kα
22
Inhibitors.

domains leading to change in protein conformation and gain in
27-32
lipid kinase activity.
Tyrosine kinases phosphorylate tyrosine residue in a panel of
different substrates and are classified as a receptor tyrosine
33
kinase (RTK) and non-receptor tyrosine kinase (NRTK).
Epidermal growth factor receptor (EGFR) is a receptor tyrosine
kinase that modulates cell signaling cascade, induces cancer cell
proliferation and growth, promotes angiogenesis and metastasis,
34-37
and causes antiapoptotic effect.
In cancer cells, the kinase
activity of EGFR is impaired by different oncogenic
mechanisms; EGFR point mutation, gene amplification, and
38
overexpression.
EGFR overexpression is observed in patients with metastatic
non-small-cell lung cancer (NSCLC) corresponding to poor
39
Fig. 2. PI3Kα pharmacophore model with 8. Aro stands for
prognosis. Mutations in the catalytic kinase domain of EGFR
G719C, L858R, and L861Q) accompany lung adenocarcinoma,
aromatic rings; Acc for H-bond acceptor; and Hyd for
(
23
40-42
hydrophobic groups. Picture made by MOE.
particularly non-small-cell lung cancer (NSCLC) , contrarily
to mutations in the extracellular domain of EGFR that cause
4
0, 42
glioblastoma.
Around 6% of breast tumors are associated
43
Indole-2-acyl chloride (6) was prepared via reaction of 1H-
indole-2-carboxylic acid (5) with excess thionyl chloride in dry
chloroform under reflux; evaporation of the solvent afforded 6
which was immediately used in the next step without further
purification. Reaction of 6 with appropriate aminoacetophenone/
benzophenone in presence of pyridine and triethylamine afforded
the corresponding caroxamides 7-10. On the other hand,
compounds 11-14 were prepared in good yields by reduction of
with EGFR overexpression. Therefore, selective targeting of
PI3Kα and/or EGFR would offer a beneficial therapeutic
approach for cancer treatment. Several chemical core structures
44-50
have been designed and synthesized targeting PI3Kα
and/or
and some of these have been reported in clinical
5, 27, 53-59
5
1, 52
EGFR
1
studies.
We report herein the synthesis of novel compounds containing
an indole nucleus recruiting ligand- and structural-based drug
design strategies; these compounds, to the best of our knowledge,
have not been previously described in the literature. In addition,
their antitumor activity against an array of cancer cell lines was
investigated in vitro.
4
compounds 7-10 with sodium borohydride (NaBH ) as a
chemoselective and simple reducing agent in a polar protic
solvent under mild conditions as outlined in Scheme 1. Structures
of the newly prepared compounds were determined with the aid
1
of various spectroscopic techniques such as FT-IR and NMR ( H
13
and C), and mass spectrometry and by elemental analysis.
O
O
O
3
CH (Ph)
HN
(ii)
(i)
C
N
OH
N
Cl
N
HN
O
5
H
6
H
H
7-10
(
iii)
O
O
F
O
Products Position
H
N
O
7,11
,12
9,13
0,14
3/CH3
O
CH (P
3
8
4/CH3
h)
N
N
3/C H5
N
H
HN
OH
6
1
4/C H5
6
H
11-14
2
1
Scheme 1. Reactions and conditions: (i) SOCl
h; (ii) Ar-NH , base, CHCl
CH OH, r.t, 6 h.
2
, CHCl
3
, reflux,
6
2
3 4
, reflux, 24 h; (iii) NaBH , dry
O
H
N
3
O
O
NH
HN
In order to examine the antiproliferative activities induced by the
newly synthesized compounds 7-14 against proteins having
kinase domain, we examined the growth inhibition in the colon
(HCT-116) and breast cancer (MDA231) cell lines both
harboring kinase domains as well as leukemia (K562) cell line as
negative control as shown in Table 1. The malignant human
colon carcinoma cell line (HCT116) contains both wild-type
OH
O
N
N
H
N
3
4
H
(
WT) and mutant (MUT) (H1047R) PI3Kα produced from a
60
primary tumor tissue culture. The metastatic human breast
cancer cell line (MDA-231) was employed as a model for
estrogen receptor negative breast cancer; it amplifies and/or
overexpresses the gene encoding EGFR resulting in poor clinical
Fig. 1. Chemical structures of some indole-based derivatives.
61-68
prognosis and high recurrence rate.
The human myeloid leukemia cell line (K562) was derived from
the pleural fluid of a patient with chronic myeloid leukemia in
69
blast crisis. The blast cells in K562 are rich in glycophorin and
they might be stimulated to produce fetal and embryonic

7
0
hemoglobin in the presence of hemin. K562 cell line represents
the primary source of chronic myeloid leukemia cells showing
significant indication of malignancy for clinical and experimental
assays to assess activators and inhibitors of the “death cascade.”
To determine the apoptotic effects of the compounds under
investigation, cells were incubated with 120 μM of each
compound for 48 hours and changes in caspase-3 enzyme
activities were evaluated. Results indicate a significant increase
in caspase-3 activity in response to 48 hours treatment for each
compound (Fig.4). Interestingly, the ability of the examined
compounds to augment apoptosis was closely relevant to their
potency as measured by their IC50 values. Compound 13 was the
most potent against the two tested cell lines and also appeared to
enhance apoptosis more significantly when compared to others.
69
studies. Additionally, no overexpression of tyrosine kinase (Bcr
7
1
-
Abl) was observed in K562 ; thus k562 is considered as a
negative control in our experiment. The prospective compounds
-14 inhibited HCT116 and MDA231 cell lines, both containing
kinase domain. Comparably no inhibition was observed in K562
cell line. This provides a proof that this scaffold targets the
kinase domain. Compound 13 exerted high potency in both
HCT116 and MDA231 cell lines.
7
Table 1. Growth inhibition IC50 (μM) after 24 hours exposure
time. Standard deviation (SD) never exceeded 5%.
IC50
HCT116
μM) ± SD
64±3
61±5
55±3
IC50
MDA 231
(μM) ± SD
57±2
IC50
K562
(μM) ± SD
>100
Compound
(
7
8
9
50±4
45±2
>100
>100
1
1
1
1
1
0
1
2
3
4
54±2
38±2
37±4
19±1
49±3
34±2
35±2
15±1
>100
>100
>100
>100
44±5
38±4
>100
Fig. 3. PI3Kα pharmacophore model with 13. Aro stands for
aromatic rings; Acc for H-bond acceptor; and Hyd for
hydrophobic groups. Picture made by MOE.
It's worth noting that attaching a methyl or phenyl moiety on
either positions 3 or 4 of the aromatic ring of compounds 7-10,
showed similar manner of inhibition in HCT116 and MDA231.
This infers that the methyl or phenyl group provided hydrophobic
interaction with the backbone of the binding site regardless of
steric effect. Interestingly, a better activity was exerted for their
structural analogues having an (OH) group instead of (CO) for
23
11-14; the IC50s dropped to half of those of 7-10. This might be
due to presence of an OH group that furnishes H-bond donor and
acceptor whereas the ketone group provides only H-bond
acceptor.
Remarkably, compounds 11 and 12 exhibited comparable
inhibition in HCT116 and MDA231. This implies that tailoring
the phenyl moiety with hydroxyethyl motif on positions 3 or 4
exerts similar activity against the kinase domain. Surprisingly,
the hydroxyphenyl motif on the 3-position of the aromatic ring,
as in 13, appeared to be the most active. On the other hand, 14
showed lower activities compared to 13 which suggests that
tailoring the 4-position is not favored. Moreover, tailoring the 3-
position is favored for the hydroxyphenyl motif. This finding
recommends that position 3 is favored for functionalities bearing
H-bond acceptor. Similarly, comparison between 11 and 13
infers that the aromatic ring might be involved in aromatic (π-π)
stacking and/or orientation in the binding cleft. Altogether, H-
bond acceptor and aromatic ring representing hydroxyphenyl
motif are essential for activity. The PI3Kα pharmacophore model
with 13 agrees with biological data and declares that core
structure of 13 fits the finger print of PI3Kα inhibitor (Fig.3).
Moreover, the absence or low activity of this scaffold against
K562 provides another clue for the selectivity of this core
nucleus towards kinases. K562 harbors esterase, lipid, and acid
Fig.4. Effects of compounds treatment on caspase-3 activity in
HCT116 and MDA231 cells after 48 hrs. Results are the means
of two independent experiments. P < 0.05 was considered
significant.
To explain the anticancer activity of the synthesized compounds
against human colon carcinoma (HCT-116) and breast cancer
(
MDA231) cell lines, we employed the crystal structures of
7
7
7
8
PI3Kα (PDB ID: 2RD0)
and EGFR (PDB ID: 3W32)
to
identify the binding interactions of these compounds in PI3Kα
and EGFR kinase domains. The binding site of 2RD0 is
composed of M772, K776, W780, I800, K802, L807, D810,
Y836, I848, E849, V850, V851, S854, T856, Q859, M922, F930,
I932, and D933. Interestingly, similar pattern of binding residues
encloses the kinase domain of 3W32: L718, G719, S720, V726,
A743, K745, M766, C775, R776, L777, L788, I789, Q791, L792,
M793, G796, C797, L844, I853, T854, D855, F856, L858, F997,
and L1001. It's worth noting that the hydrophobic and polar
residues occupy the binding clefts. Furthermore, the hydrophilic
and hydrophobic exposed surfaces of the co-crystallized ligands
are in agreement with the surrounding residues (Fig.5). The
aromatic residues furnish π-stacking aromatic interaction,
72-74
phosphatase
and thus it is considered as a negative control.
In order to determine whether growth inhibition of colon and
breast cancer cell lines was mediated by an apoptotic pathway,
we measured the caspase-3 enzyme activity using the caspase-3
75, 76
colorimetric assay. Activation of the caspase-3 pathway
is a
strong indication of apoptosis and can be employed in cellular

hydrophobic lining provides hydrophobic (van der Waals)
interaction, and polar residues offer water accessible surface area
through hydrogen-bonding, ion-dipole, and dipole-dipole
interactions. Additionally, the polar acidic or basic residues
afford an ionic (electrostatic) bonding.
accommodate the kinase domains of PI3Kα and EGFR and
approved that their binding conformations superpose the co-
crystallized docked pose (Fig. 6). The backbones of the prepared
compounds form H-bonds with E849, V851, and S854 of 2RD0
(Table 2). Remarkably, these compounds tend to form H-bonds
with M793, T854, and D855 of 3W32 (Fig. 7). The significance
of these residues and their interaction model has been reported in
14, 16-18, 82
77, 78
other computational
and experimental studies.
Fig.6. The kinase domains of (A) 2RD0 and (B) 3W32
accommodate the docked pose of the co-crystallized ligands
represented in red color and some verified molecules represented
in purple, pink, green, and blue colors. Some of key binding
residues are shown for clarity purpose.
Table 2. The Glide docking scores (Kcal/mol) of the synthesized
compounds against PI3Kα (2RD0) and EGFR (3W32). NA
means (not available).
CPD#
docking
score
Binding
residues
docking
score
Binding
residues
Fig. 5. The binding cleft of (A) 2RD0 and (B) 3W32 having co-
crystallized ligands. Green colored sphere stands for hydrophobic
residue, pink colored spheres for hydrophilic (polar) residue, red
contour circle represents acidic residue, and blue contour circle
represents basic residues.
Kcal/mol
Kcal/mol
(EGFR)
-7.91
-7.91
-7.70
-7.94
-8.05
-7.99
-9.21
(
PI3Kα)
-6.89
-7.13
-7.71
-8.13
-6.56
-6.81
-7.61
-8.36
7
8
9
V851,S854
V851,S854
V851,S854
V851,S854
E849, V851
V851,S854
V851,S854
V851,S854
M793, T854
M793, T854
NA
NA
54
M793, T854
1
1
1
0
1
2
To determine the structural-basis of binding of tested compounds
in the kinase domains of PI3Kα and EGFR, we employed Glide
7
9-81
13
D855
M793
docking
against the kinase clefts of 2RD0 and 3W32. Our
14
-8.64
Glide docking data showed that the synthesized molecules

identify the native poses in crystal structures and can successfully
predict the ligand-binding conformation.
Fig. 8. The superposition of the Glide-docked W32 pose and
its native conformation in 3W32. The native coordinates are
pink colored and the docked pose is blue. Picture made by
PYMOL.
In conclusion, a new class of PI3Kα and EGFR inhibitors having
an indole core structure has been identified via recruiting ligand-
and structure-based drug design approaches. A series of indole-2-
carboxamides have been synthesized and screened in vitro
against a panel of cancer cell lines: human colon carcinoma
(HCT-116), breast cancer (MDA231), and leukemia (K562).
Molecular docking illustrated that the synthesized compounds fit
PI3Kα and EGFR kinase domains and tend to form H-bonds with
key binding residues. Furthermore, comparable binding affinity
was observed for kinase harboring proteins (PI3Kα and EGFR).
Our biological results showed that none of the synthesized
molecules exerted any inhibitory activity in K562. However,
compound 13 exhibited high potency in HCT116 and MDA231
with IC50 values of 19 and 15 µM, respectively. Our findings
recommend that further optimization of this series would be
beneficial for colon and breast cancer treatment.
Fig. 7. Binding conformation of 8 in the kinase domains of (A)
RD0 and (B) 3W32. H-Bond is depicted in blue dotted line. H
atoms are hidden for clarity purpose. Picture made by PYMOL.
2
Docking study reveals that compounds 7-14 exerted comparable
binding affinity in the kinase domain of PI3Kα and EGFR. A
difference of 1 Kcal/mol in the docking score values against
PI3Kα and EGFR implies that these compounds have a tendency
toward the kinase domain. Interestingly, the biological data are in
agreement with molecular docking and suggest that this scaffold
might be a promising kinase inhibitor. Supplementary Fig. 1S
shows a positive relationship between the IC50s and Glide
Supplementary Material
2
2
docking scores against PI3Kα (r =0.88) and EGFR (r = 0.65).
Furthermore, the more negative binding scores of 7-14 toward
PI3Kα and EGFR infer that this scaffold might serve as a lead for
kinase inhibitors.
Detailed experimental methods, computational approaches,
spectroscopic characterization results, as well as additional
figures are listed in the supporting information and are available
at http://www.elsevier.com
Notably, the backbones of this scaffold form H-bonds with S854
of PI3Kα. We have reported previously the significance of this
14
Acknowledgments
residue to target selectively mutant H1047R PI3Kα.
This
finding prompted us to screen 7-14 against a panel of kinases and
knockout either allele of human colon cancer cell line to produce
only one specific gene in HCT116 to better discuss results based
on molecular level.
The authors wish to express their sincere appreciation to the
Hamdi Mango Center for Scientific Research at the University of
Jordan and Faculty of Pharmacy at Al-Zaytoonah University of
Jordan for their financial supports
Author Information
To evaluate the performance of Glide program, we compare the
docked pose of W32 in EGFR (PDB ID: 3W32) to its native
conformation in the crystal structure. Fig. 8 shows the
superposition of the Glide-generated W32 pose and the native
conformation in 3W32.The RMSD for heavy atoms of W32
between Glide-generated docked poses and the native poses was
Corresponding Author: *Department of Pharmacy, Faculty of
Pharmacy, Al-Zaytoonah University of Jordan, P.O. Box 130
Amman 11733 Jordan. Tel: +96264291511; Ext.: 296; Fax:
+
96264291432; E-mail address (work): dima.sabbah@zuj.edu.jo;
(home): dima_sabbah@yahoo.com
0.967 Å. Results indicate that Glide docking is capable to

*
1
+
Department of Chemistry, the University of Jordan, Amman
1942, Jordan, Tel: +96265355000; Ext: 22160; Fax:
96265355522; E-mail address (work): k.sweidan@ju.edu.jo;
34. Arora, A.; Scholar, E. M. J. Pharmacol. Exp. Ther. 2005 315, 971.
3
3
3
3
3
2
4
5. Gazdar, A. F.; Minna, J. D. PLoS Med. 2005, 2, e377.
6. Gazdar, A. F.; Minna, J. D. Cancer Prev. Res. 2008, 1, 156.
7. Gazdar, A. F. Oncogene 2009, 28, S24.
8. Ciardello, F.; Tortora, G. N. Engl. J. Med. 2008, 358, 1160.
9. Sharma, S. V.; Bell, D. W.; Settleman, J.; Haber, D. A. Nat. Rev. Cancer
007, 7, 169.
(home): kamal_sweidan@hotmail.com
0. Lynch, T. J.; Bell, D. W.; Sordella, R.; Gurubhagavatula, S.; Okimoto, R.
References and notes
A.; Brannigan, B. W. N. Engl. J. Med. 2004, 350, 2129.
4
7
4
4
1. Kobayashi, S.; Boggon, T. J.; Dayaram, T. N. Engl. J. Med. 2005, 352,
86.
2. Siegelin, M. D.; Borczuk, A. C. Lab. Invest. 2014, 94, 129.
3. Bhargava, R.; Gerald, W. L.; Li, A. R.; Pan, Q.; Lal, P.; Ladanyi, M.;
1
. La Regina, G.; Coluccia, A.; Brancale, A.; Piscitelli, F.; Gatti, V.; Maga,
G.; Samuele, A.; Pannecouque, C.; Schols, D.; Balzarini, J.; Novellino, E.;
Silvestri, R. J. Med. Chem. 2011, 54, 1587.
Chen, B. Mod. Pathol. 2005, 18, 1027.
4. Hayakawa, M.; Kaizawa, H.; Moritomo, H.; Koizumi, T.; Ohishi, T.;
2
. Pécheur, E.-I.; Lavillette, D.; Alcaras, F.; Molle, J.; Boriskin, Y. S.;
4
Roberts, M.; Cosset, F.-L.; Polyak, S. J. Biochemistry 2007, 46, 6050.
Okada, M.; Ohta, M.; Tsukamoto, S.-i.; Parker, P.; Workman, P.; Waterfield,
M. Bioorg. Med. Chem. 2006, 14, 6847.
3
2
4
. Zhao, C.; Zhao, Y.; Chai, H.; Gong, P. Bioorg. Med. Chem. 2006 14,
552.
. Biswal, S.; Sahoo, U.; Sethy, S.; Kumar, H. K. S.; Banerjee, M. Asian J.
4
5. Hayakawa, M.; Kaizawa, H.; Moritomo, H.; Koizumi, T.; Ohishi, T.;
Yamano, M.; Okada, M.; Ohta, M.; Tsukamoto, S.; Raynaud, F. I.; Workman,
P.; Waterfield, M. D.; Parker, P. Bioorg. Med. Chem. Lett. 2007, 17, 2438.
Pharm. Clin. Res. 2012, 5, 1.
. Shattat, G.; Al-Qirim, T.; Sheikha, G. A.; Al-Hiari, Y.; Sweidan, K.; Al-
5
4
6. Hayakawa, M.; Kawaguchi, K.-I.; Kaizawa, H.; Tomonobu, K.; Ohishi,
Qirim, R.; Hikmat, S.; Hamadneh, L.; Al-Kouz, S. J. Enzyme Inhib. Med.
Chem. 2013, 28, 863.
T.; Yamano, M.; Okada, M.; Ohta, M.; Tsukamoto, S.-i.; Raynaud, F. I.;
Parker, P.; Workman, P.; Waterfield, M. D. Bioorg. Med. Chem. 2007, 15,
6
8
7
. Thirumurugan, P.; Mahalaxmi, S.; Perumal, P. J. Chem. Sci. 2010, 122,
19.
. Guerra, A. S.; D.j., M.; Laranjeira, L. P.; Maia, M. B.; Colaco, N. C.; de
5
4
837.
7. Raynaud, F. I.; Eccles, S.; Clarke, P. A.; Hayes, A.; Nutley, B.; Alix, S.;
Henley, A.; Di-Stefano, F.; Ahmad, Z.; Guillard, S.; Bjerke, L. M.; Kelland,
L.; Valenti, M.; Patterson, L.; Gowan, S.; Brandon, A. d. H.; Hayakawa, M.;
Kaizawa, H.; Koizumi, T.; Ohishi, T.; Patel, S.; Saghir, N.; Parker, P.;
Waterfield, M.; Workman, P. Cancer Res. 2007, 67, 5840.
Lima, M. C.; Galdino, S. L.; Pitta, I. R.; Goncalves-Silva, T. Int.
Immunopharmacol. 2011, 11, 1816.
8
. Kaushik, N.; Kaushik, N.; Attri, P.; Kumar, N.; Kim, C.; Verma, A.; Choi,
E. Molecules 2013, 18, 6620.
4
8. Kendall, J. D.; Rewcastle, G. W.; Frederick, R.; Mawson, C.; Denny, W.
9
1
. Biersack, B.; Schobert, R. Curr. Drug Targets 2012, 13, 1705.
0. Ahmad, A.; Biersack, B.; Li, Y.; Kong, D.; Bao, B.; Schobert, R.; Padhye,
A.; Marshall, E. S.; Baguley, B. C.; Chaussade, C.; Jackson, S. P.; Shepherd,
P. R. Bioorg. Med. Chem. 2007, 15, 7677.
S. B.; Sarkar, F. H. Anticancer Agents Med. Chem. 2013, 13, 1002.
1. Liou, J. P.; Wu, Z. Y.; Kuo, C. C.; Chang, C. Y.; Lu, P. Y.; Chen, C. M.;
Hsieh, H. P.; Chang, J. Y. J. Med. Chem. 2008, 51, 4351.
2. Kuo, C. C.; Hsieh, H. P.; Pan, W. Y.; Chen, C. P.; Liou, J. P.; Lee, S. J.;
Chang, Y. L.; Chen, L. T.; Chen, C. T.; Chang, J. Y. Cancer Res. 2004, 64,
4
9. Knight, S. D.; Adams, N. D.; Burgess, J. L.; Chaudhari, A. M.; Darcy, M.
1
G.; Donatelli, C. A.; Luengo, J. I.; Newlander, K. A.; Parrish, C. A.; Ridgers,
L. H.; Sarpong, M. A.; Schmidt, S. J.; Van Aller, G. S.; Carson, J. D.;
Diamond, M. A.; Elkins, P. A.; Gardiner, C. M.; Garver, E.; Gilbert, S. A.;
Gontarek, R. R.; Jackson, J. R.; Kershner, K. L.; Luo, L.; Raha, K.; Sherk, C.
S.; Sung, C.-M.; Sutton, D.; Tummino, P. J.; Wegrzyn, R. J.; Auger, K. R.;
Dhanak, D. ACS Med. Chem. Lett. 2010, 1, 39.
1
4
1
1
5
1
5
1
621.
3. Singh, P.; Kaur, M.; Verma, P. Bioorg. Med. Chem. Lett. 2009, 19, 3054.
4. Sabbah, D. A.; Vennerstrom, J. L.; Zhong, H. J. Chem. Inf. Model. 2010,
0, 1887.
5. Sabbah, D. A.; Brattain, M. G.; Zhong, H. Curr. Med. Chem. 2011, 18,
528.
5
0. Heffron, T. P.; Wei, B.; Olivero, A.; Staben, S. T.; Tsui, V.; Do, S.;
Dotson, J.; Folkes, A. J.; Goldsmith, P.; Goldsmith, R.; Gunzner, J.; Lesnick,
J.; Lewis, C.; Mathieu, S.; Nonomiya, J.; Shuttleworth, S.; Sutherlin, D. P.;
Wan, N. C.; Wang, S.; Wiesmann, C.; Zhu, B. Y. J. Med. Chem. 2011, 54,
6. Sabbah, D. A.; Simms, N. A.; Brattain, M. G.; Vennerstrom, J. L.; Zhong,
7
5
815.
H. Bioorg. Med. Chem. Lett. 2012, 22, 876.
7. Sabbah, D. A.; Simms, N. A.; Wang, W.; Dong, Y.; Ezell, E. L.; Brattain,
M. G.; Vennerstrom, J. L.; Zhong, H. A. Bioorg. Med. Chem. 2012, 20,
1. Li, D.-D.; Fang, F.; Li, J.-R.; Du, Q.-R.; Sun, J.; Gong, H.-B.; Zhu, H.-L.
1
Bioorg. Med. Chem. Lett. 2012, 22, 5870.
2. Mao, Y.; Zhu, W.; Kong, X.; Wang, Z.; Xie, H.; Ding, J.; Terrett, N. K.;
Shen, J.; Shen, J. Bioorg. Med. Chem. 2013, 21, 3090.
5
7
1
2
1
175.
8. Sabbah, D. A.; Vennerstrom, J. L.; Zhong, H. A. J. Chem. Inf. Model.
012, 52, 3213.
9. Sabbah, D. A.; Saada, M.; Khalaf, R. A.; Bardaweel, S.; Sweidan, K.; Al-
5
5
3. Wu, P.; Hu, Y. Z. Curr. Med. Chem. 2010, 17, 4326.
4. Rodon, J.; Dienstmann, R.; Serra, V.; Tabernero, J. Nat. Rev. Clin.
Oncol. 2013, 10, 143.
Qirim, T.; Al-Zughier, A.; Halim, H. A.; Sheikha, G. A. Bioorg. Med. Chem.
Lett. 2015, 25, 3120.
5
5
4
5
5
5
6
5. Lacouture, M. E. Nat. Rev. Cancer 2006, 6, 803.
6. Wheeler, D. L.; Dunn, E. F.; Harari, P. M. Nat. Rev. Clin. Oncol. 2010, 7,
93.
7. Herbst, R. S. Int. J. Radiat. Oncol. Biol. Phys. 2004, 59, S21.
8. Sridhar, S. S.; Seymour, L.; Shepherd, F. A. Lancet Oncol. 2003, 4, 397.
9. Hynes, N. E.; Lane, H. A. Nat. Rev. Cancer 2005, 5, 341.
0. Brattain, M. G.; Levine, A. E.; Chakrabarty, S.; Yeoman, L. C.; Willson,
2
2
0. Sabbah, D. A.; Hu, J.; Zhong, H. A. Curr. Top. Med. Chem. 2016, 16, 1.
1. Sweidan, K.; Engelmann, J.; Abu Rayyan, W.; Sabbah, D.; Abu Zarga,
M.; Al-Qirim, T.; Al-Hiari, Y.; Abu Sheikha, G.; Shattat, G. Lett. Drug Des.
Discov. 2015, 12, 417.
2
2. Sweidan, K.; Sabbah, D. A.; Engelmann, J.; Halim, H. A.; Sheikha, G. A.
Lett. Drug Des. Discov. 2015, 12, 856.
3. The Molecular operating Environment Chemical Computing Group, Inc
Montreal, Quebec Canada, 2012.
J. K. V.; Long, B. Cancer Metastasis Rev. 1984, 3, 177.
2
6
6
1. R., K. C.; Kraus, M. H.; Aaronson, S. A. Science 1985, 229, 974.
2. Yokota, J.; Yamamoto, T.; Toyoshima, K.; Terada, M.; Sugimura, T.;
2
2
4. Vanhaesebroeck, B.; Waterfield, M. D. Exp. Cell. Res. 1999, 253, 239.
5. Vanhaesebroeck, B.; Guillermet-Guibert, J.; Graupera, M.; Bilanges, B.
Battifora, H.; Cline, M. J. Lancet. 1986, 1, 765.
3. Slamon, D. J.; Clark, G. M.; Wong, S. G.; Levin, W. J.; Ullrich, A.;
McGuire, W. L. Science. 1987, 235, 177.
6
Nat. Rev. Mol. Cell Biol. 2010, 11, 329.
2
2
2
2
6. Zhao, L.; Vogt, P. K. Proc. Natl. Acad. Sci. U.S.A. 2008, 105, 2652.
7. Kang, S. Y.; Bader, A. G.; Vogt, P. K. Proc. Natl. Acad. Sci. U.S.A.
005, 102, 802.
6
1
6
6
4. Price, D. J.; Grove, J. R.; Calvo, V.; Avruch, J.; Bierer, B. E. Science
992, 257, 973.
5. Hermanto, U.; Zong, C. S.; Wang, L. H. Oncogene 2001, 20, 7551.
6. Meric, F.; Lee, W. P.; Sahin, A.; Zhang, H.; Kung, H. J.; Hung, M. C.
8. Sun, M.; Hillmann, P.; Hofmann, B. T.; Hart, J. R.; Vogt, P. K. Proc.
Natl. Acad. Sci. U.S.A. 2010, 107, 15547.
9. Benvenuti, S.; Frattini, M.; Arena, S.; Zanon, C.; Cappelletti, V.;
Clin. Cancer Res. 2002, 8, 361.
7. Subik, K.; Lee, J.-F.; Baxter, L.; Strzepek, T.; Costello, D.; Crowley, P.;
Xing, L.; Hung, M.-C.; Bonfiglio, T.; Hicks, D. G.; Tang, P. Breast Cancer
2
6
Coradini, D.; Daidone, M. G.; Pilotti, S.; Pierotti, M. A.; Bardelli, A. Hum.
Mutat. 2008, 29, 284.
2
6
6
7
7
010, 4, 35.
3
3
0. Liu, Z. N.; Roberts, T. M. Cell Cycle 2006, 5, 675.
1. Miled, N.; Yan, Y.; Hon, W.-C.; Perisic, O.; Zvelebil, M.; Inbar, Y.;
8. Holliday, D. L.; Speirs, V. Breast Cancer Res. 2011, 13, 215.
9. Lozzio, C. B.; Lozzio, B. B. Blood 1975, 45, 321.
0. Koeffler, H. P.; Golde, D. W. Blood 1980, 56, 344.
1. Mahon, F. X.; Deininger, M. W. N.; Schultheis, B.; Chabrol, J.; Reiffers,
Schneidman-Duhovny, D.; Wolfson, H. J.; Backer, J. M.; Williams, R. L.
Science 2007, 317, 239.
3
2. Mandelker, D.; Gabelli, S. B.; Schmidt-Kittler, O.; Zhu, J.; Cheong, I.;
J.; Goldman, J. M.; Melo, J. V. Blood 2000, 96, 1070.
2. Klein, E. F.; Ben-Bassat, H.; Neumann, H.; Ralph, P.; Zeuthen, J.;
Polliack, A.; Vanky, F. Int. J. Cancer 1976, 18, 421.
Huang, C.-H.; Kinzler, K. W.; Vogelstein, B.; Amzel, L. M. Proc. Natl.
Acad. Sci. U.S.A. 2009, 106, 16996.
7
3
3. Paul, M. K.; Mukhopadhyay, A. K. Int. J. Med. Sci. 2004, 1, 101.

7
1
7
7
1
7
7
3. Andersson, L. C.; Nilsson, K.; Gahmberg, C. G. Int. J. Cancer 1979, 23,
43.
4. Levenson, A. S.; Jordan, V. C. Cancer Res. 1997, 57, 3071.
5. Janicke, R. U.; Sprengart, M. L.; Wati, M. R.; Porter, A. G. J. Biol. Med.
998, 273, 9357.
6. Porter, A. G.; Janicke, R. U. Cell Death Differ. 1999, 6, 99.
7. Huang, C.-H.; Mandelker, D.; Schmidt-Kittler, O.; Samuels, Y.;
Velculescu, V. E.; Kinzler, K. W.; Vogelstein, B.; Gabelli, S. B.; Amzel, L.
M. Science 2007, 318, 1744.
7
8. Kawakita, Y.; Seto, M.; Ohashi, T.; Tamura, T.; Yusa, T.; Miki, H.;
Iwata, H.; Kamiguchi, H.; Tanaka, T.; Sogabe, S.; Ohta, Y.; Ishikawa, T.
Bioorg. Med. Chem. 2013, 21, 2250.
7
9. Protein Preparation Wizard, Maestro, MacroModel, Phase, Induced Fit,
Jaguar, and Glide; Schrödinger, LLC: Portland, OR, 2012.
0. Friesner, R. A.; Banks, J. L.; Murphy, R. B.; Halgren, T. A.; Klicic, J. J.;
8
Mainz, D. T.; Repasky, M. P.; Knoll, E. H.; Shelley, M.; Perry, J. K.; Shaw,
D. E.; Francis, P.; Shenkin, P. S. J. Med. Chem. 2004, 47, 1739.
8
1. Friesner, R. A.; Murphy, R. B.; Repasky, M. P.; Frye, L. L.; Greenwood,
J. R.; Halgren, T. A.; Sanschagrin, P. C.; Mainz, D. T. J. Med. Chem. 2006,
4
8
9, 6177.
2. Zhong, H.; Tran, L. F.; Stang, J. L. J. Mol. Graph. Model. 2009, 28, 336.
.