Design, synthesis and biological evaluation of 5-amino-4-(1H-benzoimidazol-2-yl)-phenyl-1,2-dihydro-pyrrol-3-ones as inhibitors of protein kinase FGFR1

Article abstract of DOI:10.1016/j.bmc.2016.03.036

Fibroblast growth factor receptor 1 (FGFR1) plays an important role in tumorigenesis and is therefore an attractive target for anticancer therapy. Using molecular docking approach we have identified inhibitor of FGFR1 belonging to 5-amino-4-(1H-benzoimidazol-2-yl)-phenyl-1,2-dihydro-pyrrol-3-ones with IC₅₀ value of 3.5 μM. A series of derivatives of this chemical scaffold has been synthesized and evaluated for inhibition of FGFR1 kinase activity. It was revealed that the most promising compounds 5-amino-1-(3-hydroxy-phenyl)-4-(6-methyl-1H-benzoimidazol-2-yl)-1,2-dihydro-pyrrol-3-one and 5-amino-4-(1H-benzoimidazol-2-yl)-1-(3-hydroxy-phenyl)-1,2-dihydro-pyrrol-3-one inhibit FGFR1 with IC₅₀ values of 0.63 and 0.32 μM, respectively, and posses antiproliferative activity against KG1 myeloma cell line with IC₅₀ values of 5.6 and 9.3 μM. Structure-activity relationships have been studied and binding mode of this chemical class has been proposed.

Full text of DOI:10.1016/j.bmc.2016.03.036

Accepted Manuscript
Design, synthesis and biological evaluation of 5-amino-4-(1H-benzoimida-
zol-2-yl)-phenyl-1,2-dihydro-pyrrol-3-ones as inhibitors of protein kinase
FGFR1
A.A. Gryshchenko, S.S. Tarnavskiy, K.V. Levchenko, V.G. Bdzhola, G.P.
Volynets, A.G. Golub, T.P. Ruban, K.V. Vygranenko, L.L. Lukash, S.M.
Yarmoluk
PII:
S0968-0896(16)30192-4
DOI:
http://dx.doi.org/10.1016/j.bmc.2016.03.036
BMC 12885
Reference:
Bioorganic & Medicinal Chemistry
To appear in:
Received Date:
Revised Date:
Accepted Date:
18 December 2015
18 March 2016
19 March 2016
Please cite this article as: Gryshchenko, A.A., Tarnavskiy, S.S., Levchenko, K.V., Bdzhola, V.G., Volynets, G.P.,
Golub, A.G., Ruban, T.P., Vygranenko, K.V., Lukash, L.L., Yarmoluk, S.M., Design, synthesis and biological
evaluation of 5-amino-4-(1H-benzoimidazol-2-yl)-phenyl-1,2-dihydro-pyrrol-3-ones as inhibitors of protein kinase
FGFR1, Bioorganic & Medicinal Chemistry (2016), doi: http://dx.doi.org/10.1016/j.bmc.2016.03.036
This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers
we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and
review of the resulting proof before it is published in its final form. Please note that during the production process
errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.

Design, synthesis and biological evaluation of 5-amino-4-(1H-benzoimidazol-
2-yl)-phenyl-1,2-dihydro-pyrrol-3-ones as inhibitors of protein kinase FGFR1
A. A. Gryshchenko,1 S. S. Tarnavskiy,1 K. V. Levchenko,1 V. G. Bdzhola,1
G.P. Volynets,1 A. G. Golub,2 T. P. Ruban,1 K.V. Vygranenko,1 L. L. Lukash,1
S. M. Yarmoluk*1
1Institute of Molecular Biology and Genetics, NAS of Ukraine, 150
Zabolotnogo St., 03680, Kyiv, Ukraine
2OTAVA Ltd., 400 Applewood Crescent, Unit 100, Vaughan, Ontario,
L4K0C3, CANADA
*Corresponding author. Phone/Fax: +38 044 522 2458. +38 044 200 0335,
E-mail: sergiy@yarmoluk.org.ua
Abstract. Fibroblast growth factor receptor 1 (FGFR1) plays an important
role in tumorigenesis and is therefore an attractive target for anticancer therapy.
Using molecular docking approach we have identified inhibitor of FGFR1
belonging to 5-amino-4-(1H-benzoimidazol-2-yl)-phenyl-1,2-dihydro-pyrrol-3-
ones with IC50 value of 3.5 µM. A series of derivatives of this chemical scaffold
has been synthesized and evaluated for inhibition of FGFR1 kinase activity. It was
revealed that the most promising compounds 5-amino-1-(3-hydroxy-phenyl)-4-(6-
methyl-1H-benzoimidazol-2-yl)-1,2-dihydro-pyrrol-3-one and 5-amino-4-(1H-
benzoimidazol-2-yl)-1-(3-hydroxy-phenyl)-1,2-dihydro-pyrrol-3-one
inhibit
FGFR1 with IC50 values of 0.63 and 0.32 µM, respectively, and posses
antiproliferative activity against KG1 myeloma cell line with IC50 values of 5.6 and
9.3 µM. Structure-activity relationships have been studied and binding mode of
this chemical class has been proposed.
Keywords: protein kinase FGFR1; inhibitor; 5-amino-4-(1H-benzoimidazol-
2-yl)-phenyl-1,2-dihydro-pyrrol-3-ones; virtual screening; kinase assay.

1. Introduction.
Fibroblast growth factor receptor 1 (FGFR1) is a receptor tyrosine kinase
which is involved in the regulation of cell growth, cell proliferation, cell
differentiation, embryonic development, angiogenesis and other important
physiological functions. FGFR1 has been suggested to be important therapeutic
target. For example, FGFR1 is correlated closely with the occurrence and
development of lung cancer.1-10 It was demonstrated that inhibition of FGFR1 by a
specific kinase inhibitor or a dominant-negative FGFR1 construct led to
significantly decreased proliferation, clonogenicity, migration, spheroid formation,
and G1 cell cycle arrest in several mesothelioma cell lines.11 FGFR1 has been
shown to be involved in the pathogenesis of oral tongue squamous cell
carcinomas,12 sinonasal cancer,13 gastric cancer,14 pancreatic cancer,15, 16 colorectal
cancer,17, 18 renal cell carcinoma,19, 20 prostate cancer,21, 22 bladder cancer23 and
acute myeloid leukemia.24 Recently, it was reported, that oncogenic mutations of
FGFR1 occurred in a subset of patients with pilocytic astrocytoma with worse
outcome.25 Experimental evidences suggest that upregulated FGFR1 expression
appears to be associated with parathyroid carcinoma in hyperparathyroidism-jaw
tumor (HPT-JT) syndrome.26 High FGFR1 amplification is a frequent oncogenic
alteration and an independent poor prognostic factor in resected esophageal
squamous cell carcinoma.27 The integrative genomic and transcriptomic approach
indicated that FGFR1 amplification, as well as the consequent up-regulation of the
protein products, plays an important role in the aetiology of uterine leiomyomas.28
FGFR1 might be involved in endometriosis development, which could possibly
serve as a novel therapeutic target and prognostic marker for this disease.29 The
activity of FGFR1, and thereby progression of the tumor growth can be reduced by
administration of inhibitors. Therefore, the development of small molecule FGFR1
modulators is a significant area of medicinal chemistry research.

2. Results and Discussion.
Using high-throughput screening of in-house drug-like compound library
targeting ATP-binding site of FGFR1 kinase, we have identified hit compound
belonging to 5-amino-4-(1H-benzoimidazol-2-yl)-phenyl-1,2-dihydro-pyrrol-3-
ones (compound 1) with IC50 value of 3.5 µM (Figure 1). This compound has some
chemical similarity (Tanimoto coefficient = 0.4 accordingly to FCFP4 fingerprints)
with multitargeted RTK inhibitor TKI258 (dovitinib) (Figure 1).30 TKI258 is
potent ATP-competitive inhibitor of VEGFR2, FGFR1, PDGFR and is currently
undergoing Phase III clinical trial in patients with renal cell carcinoma, and is
undergoing Phase II clinical trial in patients with advanced breast cancer.31
Figure 1. Chemical structures of multikinase inhibitor TKI258 and
compound 1.
In order to find more effective inhibitors of FGFR1 a series of 5-amino-4-
(1H-benzoimidazol-2-yl)-phenyl-1,2-dihydro-pyrrol-3-one derivatives has been
synthesized and tested (Table 1). The compounds were synthesized accordingly to
Scheme 1.

H2N
O
N
c
N
R3
R3
b
N
Cl
N
NH2
NH2
a
N
N
H
H3C
N
H
H3C
N
H
H3C
H2N
H3C
O
R3
N
6
15,18,19,22,29
4a
2a
3a
3b
H2N
N
c
b
a
N
N
NH2
NH2
N
N
Cl
N
N
H2N
N
H
H
R3
O
H
O
N
2b
6
4b
1,7-14,16,17,20,21,23-28,30
d
H2N
N
c
b
N
Cl
N
N
R3
N
N
N
H2N
R3
N
N
O
CH3 O
CH3
4c
6
26,28
CH3
5
Scheme 1. The synthesis pathway for compounds 1, 7-30. Reagents and
conditions: (a) Ethyl cyanoacetate, 175°C, 2h; (b) chloroacetyl chloride, TEA,
dioxane, 5°C, then reflux 1h; (c) appropriate aniline, DMF, 130°C, 7-9h; (d)
dimethylsulfate, NaOH, water.
At the first stage, o-phenyldiamines (2a, b) were converted to the
benzimidazole acetonitrile intermediates (3a, b) after cyclocondensation reaction
with cyanoacetic acid ethyl ester. Then, active methylene group in benzimidazole
acetonitrile was acylated with chloroacetyl chloride. The intermediate
benzodiazole-4-chloro-3-oxobutanenitriles (4a, b, c) were used for synthesis of 5-
amino-4-(1H-benzoimidazol-2-yl)-phenyl-1,2-dihydro-pyrrol-3-ones (1, 7-30),
cyclization was achieved by amination with appropriate anilines (6).32, 33
Inhibitory activity of the synthesized compounds toward FGFR1 was
determined by measuring the levels of phosphorylation of the synthetic peptide
kinase substrate by radiolabeled Р32 –γ-ATP. IC50 values are presented in Table 1.

Table 1. Chemical structure and in vitro FGFR1 inhibition activity of the 5-
amino-4-(1H-benzoimidazol-2-yl)-phenyl-1,2-dihydro-pyrrol-3-ones
Compound R1
R2
R3
IC50,
µMa
3.5
>30
>30
>30
9.9
1
7
H
H
H
H
H
H
H
H
H
H
H
H
3`-OCH3
H
8
H
3`-CH3
4`-CH3
2`-OCH3
4`-OCH3
2`-OСН2CH3
2`,5`-diOCH3
2`,4`-diOCH3
2`-OCH3, 5`-
CH3
9
H
10
11
12
13
14
15
H
H
H
H
H
CH3
7
>30
3.28
>30
11
16
H
H
2`-OCH3, 5`-
Cl
0.6
17
18
19
20
21
22
23
24
25
26
27
H
CH3
CH3
H
H
CH3
H
H
H
H
H
H
H
H
H
H
H
H
H
H
3`-Cl
3`-Cl
3`-F
2`-Cl
2`-F
4`-Cl
4`-F
3`-CF3
4`-OH
1.12
1.85
7.3
>30
>30
28.2
27
10
2.12
27
CH3
H
4`-OH
3`-OH, 4`-
CH3
2.28
28
29
30
H
CH3
H
CH3
H
H
3`-OH
3`-OH
3`-OH
3.8
0.63
0.32
a – Values are the mean of at least two independent experiments

The SAR associated with different substituents on the phenyl ring (R3) of 5-
amino-4-(1H-benzoimidazol-2-yl)-phenyl-1,2-dihydro-pyrrol-3-one derivatives
was investigated. Compound 7 with unsubstituted phenyl group and compounds 8
and 9 with methyl group in meta and para positions were inactive toward FGFR1.
At the same time, compound 1 with methoxy group in meta position inhibits
FGFR1 with IC50 value of 3.5 µM. Compounds 10 and 11 with methoxy group in
ortho and para position, respectively, were less active than compound 1. Replacing
methoxy group with ethoxy in ortho position (12) resulted in a complete loss of
inhibitory activity (IC50 >30 µM) which indicates a low tolerance to substituent
size. Comparing with meta methoxy derivative 1, compound 17 with meta chlorine
substituent in phenyl ring was 3-fold more active FGFR1 inhibitor. On the other
hand, compounds with para and ortho chlorine (22, 20) and para, meta and ortho
fluorine (23, 19, 21) in phenyl ring have significantly lower activity than respective
methoxy derivatives. Introduction of bulkier trifluoromethyl group in meta position
of phenyl (24) led to reduction in potency comparing with meta halogenated
phenyl derivatives (17, 19), that can be explained by unfavorable steric interaction,
as it has been shown for ortho position (12 vs. 10). Thus, it can be concluded that
inhibitory activity of 5-amino-4-(1H-benzoimidazol-2-yl)-phenyl-1,2-dihydro-
pyrrol-3-one derivatives toward FGFR1 increases with introduction of halogen or
methoxy groups in meta position of phenyl ring. The same groups in ortho and
para positions generally led to lower activity in comparison with corresponding
meta derivatives.
The same SAR was observed for compounds with double substitution in
phenyl ring. The replacement of methyl group in meta position (15) with methoxy
group (13) leads to 3.3-fold higher activity (IC50 = 11 µM and 3.28 µM,
respectively). The substitution of meta methoxy group (13) with chlorine (16)
causes 5.5-fold increasing of inhibitory activity (IC50 = 0.6 µM).
Also, we have synthesized and tested a series of 5-amino-4-(1H-
benzoimidazol-2-yl)-phenyl-1,2-dihydro-pyrrol-3-one
derivatives
containing
hydroxyl group on the phenyl ring (25-30). Compound 25 with hydroxyl group at

para position of the phenyl ring was more efficient than analogs with methoxy
group or halogen (11, 22, 23). An introduction of hydroxyl group at meta position
of the phenyl ring leads to significant increasing of inhibitory activity toward
FGFR1 (compound 30, IC50 = 0.32 µM).
Finally, we have investigated effect of substitution of hydrogen atoms at N1
(R2) and C6 (R1) on benzimidazole heterocycle by methyl group. The presence of
methyl group at the N1 position reduces inhibitory activity in 5-10 fold (26 vs 25
and 28 vs 30). A possible reason for this effect is the blocking of hydrogen bond
donor group involved in interaction with FGFR1 ATP-binding site. Methyl group
at C6 atom only slightly decreases inhibitory activity (17 vs 18 and 29 vs 30).
In order to clarify the relationship between chemical structure of 5-amino-4-
(1H-benzoimidazol-2-yl)-phenyl-1,2-dihydro-pyrrol-3-one
derivatives
and
inhibitory activity toward FGFR1 we have studied complexes of described
compounds with ATP-binding site of FGFR1, obtained by molecular docking. The
binding mode of compound 30 with FGFR1 is presented in Figure 2. Accordingly
to the results of molecular modeling, dihydro-1H-pyrrol-3-one is involved in the
hydrophobic interactions with amino acid residues Leu630, Val561, Ala512 in the
adenine-binding region. Also, it was revealed that the carbonyl group of this
heterocycle forms hydrogen bond with amino group of Ala564 which located in
the hinge region.
The benzimidazole core is implicated in van der Waals contacts with
Tyr563, Leu484 and Gly567 residues in hydrophobic region II and builds
hydrogen bond with the carbonyl group of Ala564.
The R3 substituted phenyl ring in compound 30 is extended to a back pocket
(hydrophobic region I), which is comprised of the side chains of Lys514, Ile545,
Val561 and Ala640. Substituents on the phenyl ring can interact with amino acid
residues in the back pocket depth, and possibly have a significant impact on
binding affinity in described binding mode. The meta hydroxyl group of compound
30 forms hydrogen bond with carboxyl group of Asp641 side chain, which could
improve the binding ability of the compound 30 with FGFR1.

Figure 2. Binding mode of compound 30 in the active site of the CK2
catalytic subunit. Hydrogen bonds are shown by the green dotted lines.
Three compounds with submicromolar kinase inhibition activity (16, 29, 30)
were tested for antiproliferative activity against KG1 myeloma cell line. This
cancer cell line is characterized by constitutive activity of FGFR134 and is often
used for investigation of FGFR1 inhibitors cytotoxicity.35 The results of the
antiproliferative activities are summarized in Table 2. Unexpectedly, compound 16
was inactive in cell line. Also, antiproliferative activity of compound 29 was
higher than in 30 despite lower enzymatic potency. This fact can be explained by
significant role of methyl group on the benzimidazole heterocycle for membrane
permeability of investigated compounds. Also, we have tested compounds 16, 29,
30 for their cytotoxicity potential in “normal” non-tumorous cell line – HEK293.
In this case, the compounds did not have effect on cell viability.

Table 2. Antiproliferative activities of compounds 16, 29 and 30 against KG1 and
HEK293 cell lines
Compound KG1 cell
HEK293 cell
proliferation IC50, proliferation IC50,
µMa
>50
5.6
µMa
>50
>50
>50
16
29
30
9.3
a - Values are the mean of three independent experiments, all SD within ±15ꢀ%
Two the most promising compounds (29 and 30) were taken for kinase
selectivity analysis on the panel of 7 protein kinases (Table 3). The results of our
study have shown that these compounds tend to be selective inhibitors of FGFR1.
Table 3. Residual Activity of Protein Kinases (%) in the Presence of CK2
inhibitors at 10 µM Concentration.
Compound/Kinase FGFR1 Tie-2 с-MET Aurora А JNK3 CK2 ASK1
5
92.7
84.7
92.5
75.8
85.8
100.8 77.3 91.6
118 81.4 95.7
29
30
2.1
128.3
3. Conclusions
Virtual screening experiments allowed us to identify novel class of FGFR1
inhibitors, namely, 5-amino-4-(1H-benzoimidazol-2-yl)-phenyl-1,2-dihydro-
pyrrol-3-ones. A series of derivatives of this chemical scaffold has been
synthesized and evaluated as FGFR1 inhibitors. It was revealed that the most
active compounds 29 and 30 with submicromolar IC50 have hydroxyl substituent in
meta position of phenyl ring. These inhibitors posses antiproliferative activity

against KG1 myeloma cell line with IC50 values of 5.6 and 9.3 µM and can be
promising lead compounds for further optimization as anticancer agents.
4. Experimental
4.1. Synthetic Chemistry. Starting materials and solvents were purchased from
Acros Organics and were used without further purification.1H NMR spectra were
recorded on Varian Mercury VXR400, 400 MHz with TMS as an internal standard,
DMSO-d6 was used as solvent, chemical shifts were described as parts per million
(δ) and spin multiplicities are given as s (singlet), d (doublet), dd (double doublet),
t (triplet), q (quartet), or m (multiplet). All tested compounds displayed purity of
>95ꢀ%.
General procedure of synthesis of benzoimidazol-2-yl acetonitriles (3a,
3b). The mixture of appropriate o-phenylenediamine (2a, 2b) with 1.5 equivalent
of cyanoacetic acid ethyl ester was heated on an oil bath for 1-2 h at 175oC, the
evolved ethanol being allowed to escape. After the mixture has been cooled to
100oC dioxane was added. Then the reaction mixture was cooled down to room
temperature, precipitate was filtered, washed by ethanol, water and dried. Yield of
compounds was 65-70%.
The synthesis of N1-methylbenzoimidazol-2-yl acetonitrile (5). The 1.2
equivalent of dimethylsulfate was added by drop wise to solution benzimidazol-2-
yl acetonitrile (3b), 1.1 equivalent sodium hydroxide in water at 30oC for 1h. Then
the reaction mixture was cooled down to room temperature, precipitate was
filtered, washed by water and dried. Yield of compound was 78%.
General procedure of the synthesis of 2-(1H-benzoimidazol-2-il)-4-
chloro-3-oxo-butyronitriles (4a, 4b, 4c). The benzimidazol-2-yl acetonitriles (3a,
3b, 5) from previous synthesis stage were mixed with 1.1 equivalent of
triethylamine in dioxane and after pre-cooling to 5oC was added 1.1 equivalent of
chloroacetyl chloride. The reaction mixture was reflux for 1 h. Then the reaction
mixture was cooled down to room temperature, precipitate was filtered, washed
with water and dried. Yield of compounds was 70-85%.

General procedure of the synthesis of 5-amino-4-(1H-benzoimidazol-2-
yl)-phenyl-1,2-dihydro-pyrrol-3-one derivatives (1, 7-30). The 2-(1H-
benzoimidazol-2-il)-4-chloro-3-oxo-butyronitrile from previous synthesis stage
was solved in DMF with 1.5 equivalent of appropriate aniline and was heated at
130oC for 7-9 h. Then the reaction mixture was poured in tenfold water volume
excess. The precipitate was filtered and washed by isopropanol. All compounds
didn’t require additional purification. Yield of compounds was 50-90%.
5-amino-4-(1H-benzimidazol-2-yl)-1-(3-methoxyphenyl)-1,2-dihydro-
3H-pyrrol-3-one (1). Yield 72% (yellow crystal powder); m.p. 247°С. LC-MS
m/z 321 [M+H+], Rt = 2.02 min. 1H NMR (400 MHz, DMSO-d6) δ 3.84 (s, 3H),
4.39 (s, 2H), 6.93 (d, 1H, J= 8.1), 7.02 (d, 1H, J= 8.1), 7.03 (s, 1H), 7.02 (m, 2H),
7.40 (t, 1H J= 7.8), 7.72 (m, 2H), 8.67 (s, 2H), 13.95 (bs, 1H). Anal. Calc. for
C18H16N4O2: C, 67.49; H, 5.03; N, 17.49. Found: C, 67.44; H, 5.00; N, 17.46.
5-amino-4-(1H-benzimidazol-2-yl)-1-phenyl-1,2-dihydro-3H-pyrrol-3-
one (7). Yield 72% (grey crystal powder); m.p. 257°С. LC-MS m/z 291 [M+H+],
1
Rt = 2.09 min. H NMR (DMSO-d6) δ 4.27 (s, 2H), 7.01 (m, 2H), 7.23 (t, 1H J=
6.6), 7.42 (m, 1H), 7.46 (m, 5H), 8.01 (s, 1H), 8.95 (s, 1H), 11.57 (s, 1H). Anal.
Calc. for C17H14N4O: C, 70.33; H, 4.86; N,19.30%. Found: C, 70.31; H, 4.83%; N,
19.31.
5-amino-4-(1H-benzimidazol-2-yl)-1-(3-methylphenyl)-1,2-dihydro-3H-
pyrrol-3-one (8). Yield 71% (white crystal powder); m.p. 187°С. LC-MS m/z 305
[M+H+], Rt = 2.14 min. 1H NMR (DMSO-d6) δ 2.40 (s, 3H), 4.22 (s, 2H), 7.02 (m,
3H), 7.22 (d, 1H J= 7.8), 7.27 (s, 1H), 7.31 (t, 1H J= 6.8), 7.44 (m, 2H), 7.75 (bs,
1H), 9.00 (bs, 1H), 11.32 (bs, 1H). Anal. Calc. for C18H16N4O: C, 71.04; H, 5.30;
N, 18.41. Found: C, 71.01; H, 5.28; N, 18.39.
5-amino-4-(1H-benzimidazol-2-yl)-1-(4-methylphenyl)-1,2-dihydro-3H-
pyrrol-3-one (9). Yield 71% (yellow crystal powder); m.p. 292°С (dec.). LC-MS
1
m/z 305 [M+H+], Rt = 2.14 min. H NMR (DMSO-d6) δ 2.36 (s, 3H), 4.19 (s, 2H),
7.02 (m, 2H), 7.24 (d, 2H J= 7.8), 7.29 (d, 2H J= 7.8), 7.46 (m, 2H), 7.75 (s, 1H),

9.00 (s, 1H), 11.32 (s, 1H). Anal. Calc. for C18H16N4O: C, 71.04; H, 5.30; N, 18.41.
Found: C, 71.03; H, 5.29; N, 18.38.
5-amino-4-(1H-benzimidazol-2-yl)-1-(2-methoxyphenyl)-1,2-dihydro-
3H-pyrrol-3-one (10). Yield 72% (yellow crystal powder); m.p. 282°С. LC-MS
1
m/z 321 [M+H+], Rt = 2.08 min. H NMR (DMSO-d6) δ 3.89 (s, 3H), 4.09 (s, 2H),
7.01 (m, 2H), 7.06 (t, 1H J= 7.8), 7.15 (d, 1H, СН J= 8.3), 7.46 (m, 3H), 7.75 (s,
1H), 8.96 (s, 1H), 11.46 (s, 1H). Anal. Calc. for C18H16N4O2: C, 67.49; H, 5.03; N,
17.49. Found: C, 67.47; H, 5.02; N, 17.48.
5-amino-4-(1H-benzimidazol-2-yl)-1-(4-methoxyphenyl)-1,2-dihydro-
3H-pyrrol-3-one (11). Yield 72% (brown crystal powder); m.p. 257°С. LC-MS
1
m/z 321 [M+H+], Rt = 2.08 min. H NMR (DMSO-d6) δ 3.82 (s, 3H), 4.39 (s, 2H),
7.11 (d, 2H, СН J= 8.2), 7.38 (m, 2H), 7.43 (d, 2H, СН J= 8.2), 7.72 (m, 2H), 8.48
(s, 2H), 13.77 (s, 1H). Anal. Calc. for C18H16N4O2: C, 67.49; H, 5.03; N, 17.49.
Found: C, 67.48; H, 5.02; N, 17.47.
5-amino-4-(1H-benzimidazol-2-yl)-1-(2-ethoxyphenyl)-1,2-dihydro-3H-
pyrrol-3-one (12). Yield 72% (yellow crystal powder); m.p. 218°С. LC-MS m/z
1
335 [M+H+], Rt = 2.09 min. H NMR (DMSO-d6) δ 1.38 (t, 3H J= 8.3), 4.16 (m,
4H), 7.11 (m, 4H), 7.36 (m, 2H), 7.66 (m, 3H), 8.94 (s, 1H), 11.42 (s, 1H). Anal.
Calc. for C19H18N4O2: C, 68.25; H, 5.43; N, 16.76. Found: C, 68.22; H, 5.42; N,
16.74.
5-amino-4-(1H-benzimidazol-2-yl)-1-(2,5-dimethoxyphenyl)-1,2-
dihydro-3H-pyrrol-3-one (13). Yield 72% (light beige crystal powder); m.p.
1
244°С. LC-MS m/z 351 [M+H+], Rt = 2.06 min. H NMR (DMSO-d6) δ 3.89 (s,
3H), 3.92 (s, 3H), 4.16 (s, 2H), 6.89 (d, 1H, СН J= 7.8), 6.98 (s, 2H), 7.02 (d, 1H,
СН J= 7.8), 7.24(m, 2H), 7.75 (m, 2H), 9.12 (m, 2H), 8.78 (s, 1H), 14.17 (s, 1H).
Anal. Calc. for C19H18N4O3: C, 68.13; H, 5.18; N, 15.99. Found: C, 68.10; H, 5.17;
N, 15.97.
5-amino-4-(1H-benzimidazol-2-yl)-1-(2,4-dimethoxyphenyl)-1,2-
dihydro-3H-pyrrol-3-one (14). Yield 72% (yellow crystal powder); m.p. 247°С.
1
LC-MS m/z 351 [M+H+], Rt = 2.05 min. H NMR (DMSO-d6) δ 3.83 (s, 3H), 3.88

(s, 3H), 4.06 (s, 2H), 6.53 (d, 1H, СН J= 7.8), 6.59 (s, 2H), 7.08 (m, 2H), 7.23 (d,
1H, СН J= 7.8), 7.59 (m, 3H), 8.78 (s, 1H), 11.47 (s, 1H). Anal. Calc. for
C19H18N4O3: C, 68.13; H, 5.18; N, 15.99. Found: C, 68.11; H, 5.18; N, 15.96.
5-amino-1-(2-methoxy-5-methylphenyl)-4-(6-methyl-1H-benzimidazol-
2-yl)-1,2-dihydro-3H-pyrrol-3-one (15). Yield 71% (brown crystal powder); m.p.
1
242°С. LC-MS m/z 349 [M+H+], Rt = 2.14 min. H NMR (DMSO-d6) δ 2.28 (s,
3H), 2.46 (s, 3H), 3.82 (s, 3H), 4.22 (s, 2H), 7.11 (d, 2H J= 7.8 ), 7.18 (d, 2H J=
7.8), 7.27 (m, 2H), 7.49 (s, 1H), 7.57 (d, 1H J= 7.8), 8.15 (s, 1H), 8.50 (s, 1H),
11.46 (s, 1H). Anal. Calc. for C20H20N4O2: C, 68.95; H, 5.79; N, 16.08. Found: C,
68.92; H, 5.77; N, 16.06.
5-amino-4-(1H-benzimidazol-2-yl)-1-(5-chloro-2-methoxyphenyl)-1,2-
dihydro-3H-pyrrol-3-one (16). Yield 73% (yellow crystal powder); m.p. 284°С.
1
LC-MS m/z 355 [M+H+], Rt = 2.11 min. H NMR (DMSO-d6) δ 3.87 (s, 3H), 4.21
(s, 2H), 7.17 (d, 1H, СН J= 8.8), 7.31 (m, 2H), 7.42 (d, 1H, СН J= 8.8), 7.49 (s,
1H), 7.73 (m, 2H), 8.75 (s, 2H), 14.02 (s, 1H). Anal. Calc. for C18H15ClN4O2: C,
60.94; H, 4.26; Cl, 9,99; N, 15.79. Found: C, 60.90; H, 4.25; Cl, 9,97; N, 15.76.
5-amino-4-(1H-benzimidazol-2-yl)-1-(3-chlorophenyl)-1,2-dihydro-3H-
pyrrol-3-one (17). Yield 73% (grey crystal powder); m.p. 234°С. LC-MS m/z 325
[M+H+], Rt = 2.11 min. 1H NMR (DMSO-d6) δ 4.30 (s, 2H), 7.09 (m, 2H), 7.31 (d,
1H, СН J= 7.1), 7.41 (d, 1H, СН J= 7.6), 7.49 (m, 4H), 8.29 (s, 1H), 8.99 (s, 1H),
11.60 (s, 1H). Anal. Calc. for C17H13ClN4O: C, 62.87; H, 4.03; Cl, 10,92; N, 17.25.
Found: C, 62.84; H, 4.02; Cl, 10,90; N, 17.23.
5-amino-1-(3-chlorophenyl)-4-(6-methyl-1H-benzimidazol-2-yl)-1,2-
dihydro-3H-pyrrol-3-one (18). Yield 70% (grey crystal powder); m.p. 258°С.
1
LC-MS m/z 339 [M+H+], Rt = 2.16 min. H NMR (DMSO-d6) δ 2.41 (s, 3H), 4.26
(s, 2H), 6.83 (m, 1H), 7.27 (m, 3H), 7.39 (d, 1H, СН J= 7.8), 7.46 (d, 1H, СН J=
8.1), 7.50 (d, 1H, СН J= 8.1), 7.51 (s, 1H), 8.18 (s, 1H), 9.03 (s, 1H), 11.43 (s,
1H). Anal. Calc. for C18H15ClN4O: C, 63.81; H, 4.46; Cl, 10,46; N, 16.54. Found:
C, 63.80; H, 4.45; Cl, 10,44; N, 16.51.

5-amino-1-(3-fluorophenyl)-4-(6-methyl-1H-benzimidazol-2-yl)-1,2-
dihydro-3H-pyrrol-3-one (19). Yield 70% (grey crystal powder); m.p. 264°С.
1
LC-MS m/z 323 [M+H+], Rt = 2.10 min. H NMR (DMSO-d6) δ 2.42 (s, 3H), 4.28
(s, 2H), 6.86 (d, 1H, СН J= 5.9), 7.05 (t, 1H J= 7.8), 7.31 (m, 4H), 7.50 (q, 1H, СН
J= 6.8), 8.15 (s, 1H), 9.04 (s, 1H), 11.43 (s, 1H). Anal. Calc. for C18H15FN4O: C,
67.07; H, 4.69; N, 17.38. Found: C, 67.03; H, 4.68; N, 17.35.
5-amino-4-(1H-benzimidazol-2-yl)-1-(2-chlorophenyl)-1,2-dihydro-3H-
pyrrol-3-one (20). Yield 70% (white crystal powder); m.p. 269°С. LC-MS m/z
1
325 [M+H+], Rt = 2.13 min. H NMR (DMSO-d6) δ 4.18 (s, 2H), 7.22 (m, 2H),
7.47 (m, 2H), 7.62 (m, 2H), 7.73 (m, 2H), 7.50 (q, 1H, СН J= 6.8), 8.18 (s, 1H),
9.18 (s, 1H), 11.46 (s, 1H). Anal. Calc. for C17H13ClN4O: C, 62.87; H, 4.03; Cl,
10,92; N, 17.25. Found: C, 62.85; H, 4.02; Cl, 10,92; N, 17.22.
5-amino-4-(1H-benzimidazol-2-yl)-1-(2- fluorophenyl)-1,2-dihydro-3H-
pyrrol-3-one (21). Yield 70% (beige crystal powder); m.p. 263°С. LC-MS m/z
1
325 [M+H+], Rt = 2.10 min. H NMR (DMSO-d6) δ 4.22 (s, 2H), 7.32 (m, 4H),
7.48 (q, 1H, СН J= 6.8), 7.59 (t, 1H J= 7.8), 7.74 (m, 2H), 9.08 (s, 2H), 11.57 (s,
1H). Anal. Calc. for C17H13FN4O: C, 66.23; H, 4.25; N, 18.17. Found: C, 66.20; H,
4.24; N, 18.15.
5-amino-1-(4-chlorophenyl)-4-(6-methyl-1H-benzimidazol-2-yl)-1,2-
dihydro-3H-pyrrol-3-one (22). Yield 70% (beige crystal powder); m.p. 291°C.
1
LC-MS m/z 339 [M+H+], Rt = 2.15 min. H NMR (DMSO-d6) δ 2.41 (s, 3H), 4.26
(s, 2H), 6.83 (m, 1H), 7.27 (m, 3H), 7.39 (d, 1H, СН J= 7.8), 7.46 (d, 1H, СН J=
8.1), 7.50 (d, 1H, СН J= 8.1), 7.51 (s, 1H), 8.18 (s, 1H), 9.03 (s, 1H), 11.43 (s,
1H). Anal. Calc. for C18H15ClN4O: C, 63.81; H, 4.46; Cl, 10,46; N, 16.54. Found:
C, 63.79; H, 4.46; Cl, 10.43; N, 16.51.
5-amino-4-(1H-benzimidazol-2-yl)-1-(4-fluorophenyl)-1,2-dihydro-3H-
pyrrol-3-one (23). Yield 77% (yellow crystal powder); m.p. 292°С (dec.). LC-MS
1
m/z 309 [M+H+], Rt = 2.12 min. H NMR (DMSO-d6) δ 2.42 (s, 3H), 4.31 (s, 2H),
6.89 (d, 2H J= 7.8), 7.33 (s, 1H), 7.41 (d, 2H J= 7.8), 7.51 (d, 2H J= 8.3), 7.56 (d,

2H J= 8.3), 8.15 (s, 1H), 8.88 (s, 1H), 11.56 (s, 1H). Anal. Calc. for C17H13FN4O:
C, 66.23; H, 4.25; N, 18.17. Found: C, 66.21; H, 4.24; N, 18.14.
5-amino-4-(1H-benzimidazol-2-yl)-1-[3-(trifluoromethyl)phenyl]-1,2-
dihydro-3H-pyrrol-3-one (24). Yield 70% (white crystal powder); m.p. 242°С.
1
LC-MS m/z 359 [M+H+], Rt = 2.10 min. H NMR (DMSO-d6) δ 4.28 (s, 2H), 7.03
(m, 2H), 7.46 (m, 2H), 7.57 (d, 1H, СН J= 7.8), 7.64 (t, 1H, СН J= 7.6), 7.76 (s,
1H, СН), 8.17 (s, 1H), 9.08 (s, 1H), 11.43 (s, 1H). Anal. Calc. for C18H13F3N4O: C,
60.34; H, 3.66; N, 15.64. Found: C, 60.31; H, 3.65; N, 15.62.
5-amino-4-(1H-benzimidazol-2-yl)-1-(4-hydroxyphenyl)-1,2-dihydro-
3H-pyrrol-3-one (25). Yield 70% (light grey crystal powder); m.p. 238°С. LC-MS
1
m/z 307 [M+H+], Rt = 2.02 min. H NMR (DMSO-d6) δ 4.19 (s, 2H), 6.86 (d, 2H,
СН J= 7.8), 7.09 (m, 2H), 7.16 (d, 2H, СН J= 7.8), 7.48 (m, 2H), 7.50 (d, 1H, СН
J= 8.1), 7.51 (s, 1H), 8.23 (s, 1H), 9.38 (s, 1H), 11.41 (s, 1H). Anal. Calc. for
C17H14N4O2: C, 66.66; H, 4.61; N, 18.29. Found: C, 66.62; H, 4.60; N, 18.26.
5-amino-1-(4-hydroxyphenyl)-4-(1-methyl-1H-benzimidazol-2-yl)-1,2-
dihydro-3H-pyrrol-3-one (26). Yield 70% (brown crystal powder); m.p. 163°С.
1
LC-MS m/z 321 [M+H+], Rt = 2.01 min. H NMR (DMSO-d6) δ 4.02 (s, 3H), 4.12
(s, 2H), 6.86 (d, 2H, СН J= 7.8), 7.11 (m, 2H), 7.22 (d, 2H, СН J= 7.8), 7.41 (m,
1H), 7.48 (m, 1H), 7.61 (s, 1H), 8.73 (s, 1H), 9.58 (s, 1H), 11.37 (s, 1H). Calc. for
C18H16N4O2: C, 67.49; H, 5.03; N, 17.49. Found: C, 67.45; H, 5.02; N, 17.47.
5-amino-4-(1H-benzimidazol-2-yl)-1-(3-hydroxy-4-methylphenyl)-1,2-
dihydro-3H-pyrrol-3-one (27). Yield 78% (orange crystal powder); m.p.
1
>300°С(dec.). LC-MS m/z 321 [M+H+], Rt = 2.03 min. H NMR (DMSO-d6) δ
2.17 (s, 3H), 4.16 (s, 2H), 6.74 (d, 1H, СН J= 7.8), 6.82 (s, 1H), 7.01 (m, 2H), 7.11
(d, 1H, СН J= 7.8), 7.46 (m, 2H), 7.84 (s, 1H), 9.01 (s, 1H), 9.03 (s, 1H), 9.58 (s,
1H), 11.51 (s, 1H). Calc. for C18H16N4O2: C, 67.49; H, 5.03; N, 17.49. Found: C,
67.46; H, 5.03; N, 17.46.
5-amino-1-(3-hydroxyphenyl)-4-(1-methyl-1H-benzimiazol-2-yl)-1,2-
dihydro-3H-pyrrol-3-one (28). Yield 73% (brown crystal powder); m.p. 172°С.
1
LC-MS m/z 321 [M+H+], Rt = 1.73 min. H NMR (DMSO-d6) δ 4.00 (s, 3H), 4.16

(s, 2H), 6.70 (d, 1H, СН J= 6.8), 6.81 (s, 1H), 6.82 (d, 1H, СН J= 7.2), 7.11 (m,
2H), 7.25 (t, 1H J= 7.4), 7.35 (m, 1H), 7.47 (m, 1H), 8.95 (s, 2H), 9.63 (s, 1H).
Calc. for C18H16N4O2: C, 67.49; H, 5.03; N, 17.49. Found: C, 67.49; H, 5.00; N,
17.44.
5-amino-1-(3-hydroxyphenyl)-4-(6-methyl-1H-benzimidazol-2-yl)-1,2-
dihydro-3H-pyrrol-3-one (29). Yield 71% (brown crystal powder); m.p. 278°С.
1
LC-MS m/z 321 [M+H+], Rt = 1.79 min. H NMR (DMSO-d6) δ 2.37 (s, 3H), 4.29
(s, 2H), 6.73 (d, 1H, СН J= 7.2), 6.87 (m, 3H), 7.27 (d, 1H, СН J= 8.5), 7.32 (t, 1H
J= 8.3), 7.39 (d, 1H, СН J= 7.8), 8.02 (bs, 1H), 8.91 (bs, 1H), 9.85 (bs, 1H), 11.60
13
(bs, 1H). C NMR (DMSO-d6) δ 22.00, 59.43, 88.39, 111.43, 113.94, 114.63,
130.69, 139.16, 159.08, 165.73, 186.30; Calc. for C18H16N4O2: C, 67.49; H, 5.03;
N, 17.49. Found: C, 67.48; H, 5.01; N, 17.45.
5-amino-4-(1H-benzimidazol-2-yl)-1-(3-hydroxyphenyl)-1,2-dihydro-
3H-pyrrol-3-one (30). Yield 73% (rose crystal powder); m.p. 234°С. LC-MS m/z
1
307 [M+H+], Rt = 1.70 min. H NMR (DMSO-d6) δ 4.20 (s, 2H), 6.69 (d, 1H, СН
J= 7.6), 6.82 (s, 1H), 6.83 (d, 1H, СН J= 7.8), 7.00 (m, 2H), 7.24 (t, 1H J= 7.6),
7.41 (m, 1H), 7.46 (m, 1H), 7.94 (s, 1H), 8.88 (bs, 1H), 9.63 (bs, 1H), 11.53 (bs,
1H). 13C NMR (DMSO-d6) δ 59.54, 96.23, 111.51, 113.99, 114.69, 130.68, 139.11,
159.08, 165.71, 186.44; Calc. for C17H14N4O2: C, 66.66; H, 4.61; N, 18.29. Found:
C, 66.63; H, 4.60; N, 18.27.
4.2. Kinase assay. The FGFR1 kinase assays with recombinant cytoplasmic
domain of the FGFR1 tyrosine kinase (Millipore, Cat. N. 14-582) were performed
in a total volume of 30 µl containing 10 mM MOPS (рН 7.2), 0.1 mM sodium
orthovanadate, 0.2 mM EDTA, 0.002 % Brij 35, 0.2 mg/ml BSA, 0.02 % β-
mercaptoethanol, 250 µM of peptide substrate (KKKSPGEYVNIEFG, GenScript),
various concentrations of inhibitor dissolved in DMSO (final DMSO concentration
in probe less than 1%) and 10 mU of enzyme. The reaction was initiated by the
addition of ATP (50 µM ATP, 25 mM MgAc containing 0.1 µCi of [γ-32P]ATP
per probe) and samples incubated at 30°C for 25 min. The reaction was terminated
by the addition of 5% phosphoric acid and the precipitation of material onto

phosphocellulose filters “Whatman P81”. Filters were washed three times with
0.75 % phosphoric acid and the incorporation of [32P] into the peptide substrate
was determined by counting the radioactivity retained on the filters in a
PerkinElmer scintillation counter. Kinase residual activity was expressed in
percentages with respect to DMSO control. The concentration of compound that
inhibited enzymatic activity by 50% (IC50) was determined graphically.
Inhibition of selectivity panel kinases was perfomed according to enzyme
provider protocols (Millipore). ATP concentration in reaction mixture was 100
µM.
4.3. Molecular docking. Ligands for the docking were prepared with Vega ZZ.36
Docking was carried out in crystal structure of FGFR1 (pdb bank code 3GQI) by
Autodock 4.2 with default parameters.37 Docking results were analyzed with
AutoDockTools-1.5.6.37
4.4. Cellular Proliferation Assay. KG1 and HEK293 cell viability was examined
using a standard MTT assay.38 Cells were grown in DMEM containing 10% fetal
bovine serum (FBS), 100 mg/mL penicillin, and 100 mg/mL streptomycin in
humidified air at 37oC with 5% CO2. Viable cells were seeded into 96-well tissue
plate at 2X105 cells/mL preincubated for 24 hours and treated with the synthesized
compounds (compounds in DMSO solution, final DMSO concentration less than
0.5%) at various concentrations. After 72 h treatment, the cells were incubated
with 15 µl MTT (3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl-tetrazolium bromide
from Sigma) solution (5 mg/mL) for 4 h at 37oC, 5% CO2. The formazan
precipitates were dissolved in 200 µl DMSO and the absorbance at 540 nm of each
well was measured by spectrofluorometer MR 700 (Dynatech). The cell viability
was expressed as a percentage relative to the untreated control cells.
Acknowledgements.
This research was supported by grants from the National Academy of
Sciences of Ukraine (0107U003345 and 0107U004939).
References.

1. Wu, J.; Wei, T.; Tang, Q.; Weng, B.; Li, W.; Jiang, X.; Ding, T.; Li, X.; Liang,
G.; Cai, Y.; Ji, J. BMC Cancer. 2015, 15, 276.
2. Zhao, D.; Lu, Y.; Yang, C.; Zhou, X.; Xu, Z. Tumour Biol. 2015, 36, 3637.
3. Thomas, A.; Lee, J.H.; Abdullaev, Z.; Park. K.S.; Pineda, M.; Saidkhodjaeva,
L.; Miettinen, M.; Wang, Y.; Pack, S.D.; Giaccone, G. J. Thorac. Oncol. 2014, 9,
567.
4. Wang, Y.; Cai, Y.; Ji, J.; Liu, Z.; Zhao, Y.; Wei, T.; Shen, X.; Zhang, X.; Li, X.;
Liang, G. Cancer Lett. 2014, 344, 82.
5. Lockwood, W.; Politi, K. Cancer Discov. 2014, 4, 152.
6. Ma, L.; Zhang, S. Zhongguo Fei Ai Za Zhi. 2013, 16, 671.
7. Preusser, M.; Berghoff, A.S.; Berger, W.; Ilhan-Mutlu, A.; Dinhof, C.;
Widhalm, G.; Dieckmann, K.; Wöhrer, A.; Hackl, M.; von Deimling, A.; Streubel,
B.; Birner. P. Lung Cancer. 2014, 83, 83.
8. Tran, T.N.; Selinger, C.I.; Kohonen-Corish, M.R.; McCaughan, B.C.; Kennedy,
C.W.; O’Toole, S.A.; Cooper W.A. Lung Cancer. 2013, 81, 462.
9. Ren, M.; Hong, M.; Liu, G.; Wang, H.; Patel, V.; Biddinger, P.; Silva, J.;
Cowell, J.; Hao, Z. Oncol. Rep. 2013, 29, 2181.
10. Schildhaus, H.U.; Nogova, L.; Wolf, J.; Buettner, R. Transl. Lung Cancer Res.
2013, 2, 92.
11. Schelch, K.; Hoda, M.A.; Klikovits, T.; Münzker, J.; Ghanim, B.; Wagner, C.;
Garay, T.; Laszlo, V.; Setinek, U.; Dome, B.; Filipits, M.; Pirker, C.; Heffeter, P.;
Selzer, E.; Tovari, J.; Torok, S.; Kenessey, I.; Holzmann, K.; Grasl-Kraupp, B.;
Marian, B.; Klepetko, W.; Berger, W.; Hegedus, B.; Grusch, M. Am. J. Respir.
Crit. Care Med. 2014, 190, 763.
12. Young, R.J.; Lim, A.M.; Angel, C.; Collins, M.; Deb, S.; Corry, J.; Wiesenfeld,
D.; Kleid, S.; Sigston, E.; Lyons, B.; Russell, P.A.; Wright, G.; McArthur, G.A.;
Fox, S.B.; Rischin, D.; Solomon, B. Oral. Oncol. 2013, 49, 576.
13. Schröck, A.; Göke, F.; Wagner, P.; Bode, M.; Franzen, A.; Huss, S.; Agaimy,
A.; Ihrler, S.; Kirsten, R.; Kristiansen, G.; Bootz, F.; Lengerke, C., Perner, S. Head
Neck. 2014, 36, 1253.

14. Xu, C.; Li, W.; Qiu, P.; Xia, Y.; Du, X.; Wang, F.; Shen, L.; Chen, Q.; Zhao,
Y.; Jin, R.; Wu, J.; Liang, G.; Li. X. Anticancer Drugs. 2015, 26, 379.
15. Lehnen, N.C.; von Mässenhausen, A.; Kalthoff, H.; Zhou, H.; Glowka, T.;
Schütte, U.; Höller, T.; Riesner, K.; Boehm, D.; Merkelbach-Bruse, S.; Kirfel, J.;
Perner, S.; Gütgemann, I. Histopathology. 2013, 63, 157.
16. Coleman, S.J.; Chioni, A.M.; Ghallab, M.; Anderson, R.K.; Lemoine, N.R.;
Kocher, H.M.; Grose. R.P. EMBO Mol. Med. 2014, 6, 467.
17. Kwak, Y.; Nam, S.K.; Seo, A.N.; Kim, D.W.; Kang, S.B.; Kim, W.H.; Lee,
H.S. Pathobiology. 2015, 82, 76.
18. Göke, F.; Göke, A.; von Mässenhausen, A.; Franzen, A.; Sharma, R.; Kirsten,
R.; Böhm, D.; Kristiansen, G.; Stenzinger, A.; Wynes, M.; Hirsch, F.R.; Weichert,
W.; Heasley, L.; Buettner, R.; Perner, S. Digestion. 2013, 88, 172.
19. Tsimafeyeu, I.; Bratslavsky, G. Oncology. 2015, 88, 321.
20. Tsimafeyeu, I.; Zaveleva, E.; Stepanova, E.; Low, W. Invest New Drugs. 2013,
31, 1436.
21. Wan, X.; Corn, P.G.; Yang, J.; Palanisamv, N.; Starbuck, M.W.; Efstathiou, E.;
Tapia, E.M.; Zurita, A.J.; Aparicio, A.; Ravoori, M.K.; Vazquez, E.S.; Robinson,
D.R.; Wu, Y.M.; Cao, X.; Iyer, M.K.; McKeehan, W.; Kundra, V.; Wang, F.;
Troncoso, P.; Chinnaiyan, A.M.; Logothetis, C.J.; Navone, N.M. Sci. Transl. Med.
2014, 252, 252ra122.
22. Yang, F.; Zhang, Y.; Ressler, S.J.; Ittmann, M.M.; Ayala, G.E.; Dang, T.D.;
Wang, F.; Rowley, D.R. Cancer. Res. 2013, 73, 3716.
23. Cheng, T.; Roth, B.; Choi, W.; Black, P.C.; Dinney, C.; McConkey D.J. PLoS
One. 2013, 8, e57284.
24. Lee, H.; Kim, M.; Lim, J.; Kim, Y.; Han, K.; Cho, B.S.; Kim. H.J. Int. J.
Hematol. 2013, 97, 808.
25. Becker, A.P.; Scapulatempo-Neto, C.; Carloni, A.C.; Paulino, A.; Sheren, J.;
Aisner, D.L.; Musselwhite, E.; Clara, C.; Machado, H.R.; Oliveira, R.S.; Neder, L.;
Varella-Garcia, M.; Reis R.M. J. Neuropathol. Exp. Neurol. 2015, 74, 743.

26. Lee, J.Y.; Kim, S.Y.; Mo, E.Y.; Kim, E.S.; Han, J.H.; Maeng, L.S.; Lee, A.H.;
Eun, J.W.; Nam, S.W.; Moon. S.D. Int. J. Oncol. 2014, 45, 641.
27. Kim, H.S.; Lee, S.E.; Bae, Y.S.; Kim, D.J.; Lee, C.G.; Hur, J.; Chung, H.;
Park, J.C.; da Jung, H.; Shin, S.K.; Lee, S.K.; Lee, Y.C.; Kim, H.R.; Moon, Y.W.;
Kim, J.H.; Shim, Y.M.; Jewell, S.S.; Kim, H.; Choi, Y.L.; Cho. B.C. Oncotarget.
2015, 6, 2562.
28. Cirilo, P.D.; Marchi, F.A.; Barros Filho Mde, C.; Rocha, R.M.; Domingues,
M.A.; Jurisica, I.; Pontes, A.; Rogatto. S.R. PLoS One. 2013, 8, e57901.
29. Zhao, L.; Yang, H.; Xuan, Y.; Luo, Z.; Lin, Q.; Zhao, J.; Ren, N.; Zhou, S.;
Zhao, X. Eur. J. Obstet. Gynecol. Reprod. Biol. 2015, 184, 117.
30. Renhowe, P.A.; Pecchi, S.; Shafer, C.M.; Machajewski, T.D.; Jazan, E.M.;
Taylor, C.; Antonios-McCrea, W.; McBride, C.M.; Frazier, K.; Wiesmann, M.;
Lapointe, G.R.; Feucht, P.H.; Warne, R.L.; Heise, C.C.; Menezes, D.; Aardalen,
K.; Ye, H.; He, M.; Le, V.; Vora, J.; Jansen, J.M.; Wernette-Hammond, M.E.;
Harris A.L. J Med Chem. 2009, 52, 278.
31. André, F.; Bachelot, T.; Campone, M.; Dalenc, F.; Perez-Garcia, J.M.; Hurvitz,
S.A.; Turner, N.; Rugo, H.; Smith, J.W.; Deudon, S.; Shi, M.; Zhang, Y.; Kay, A.;
Porta, D.G.; Yovine, A.; Baselga J. Clin Cancer Res. 2013, 19, 3693.
32. Volovenko, Yu.M.; Tverdokhlebov, A.V.; Gorulya, A.P.; Shishkina, S.V.;
Zubatyuk, R.I.; Shishkin, O.V. Eur. J. Org. Chem. 2002, 4, 663.
33. Tverdokhlebov, A.P.; Denisenko, A.V.; Tolmachev, A.A.; Volovenko. Yu.M.
Synthesis. 2007, 12, 1811.
34. Gu, T.L.; Goss, V.L.; Reeves, C.; Popova, L.; Nardone, J.; Macneill, J.;
Walters, D.K.; Wang, Y.; Rush, J.; Comb, M.J.; Druker, B.J.; Polakiewicz, R.D.
Blood. 2006, 108, 4202.
35. Ren, M.; Qin, H.; Ren, R.; Cowell, J.K. Leukemia. 2013, 27, 32.
36. Pedretti, A.; Villa, L.; Vistoli, G. J Comput Aided Mol Des. 2004, 18, 167.
37. Morris, G.M.; Huey, R.; Lindstrom, W.; Sanner, M.F.; Belew, R.K.; Goodsell
D.S.; Olson, A.J. J. Comput. Chem. 2009, 16, 2785.
38. Mosmann, T. J. Immunol. Methods. 1983, 16, 55.

Graphical Abstract