Structure-Based Design of Tetrahydroisoquinoline-7-carboxamides as Selective Discoidin Domain Receptor 1 (DDR1) Inhibitors

Article abstract of DOI:10.1021/acs.jmedchem.6b00140

The structure-based design of 1, 2, 3, 4-tetrahydroisoquinoline derivatives as selective DDR1 inhibitors is reported. One of the representative compounds, 6j, binds to DDR1 with a K_d value of 4.7 nM and suppresses its kinase activity with an IC₅₀ value of 9.4 nM, but it is significantly less potent for a panel of 400 nonmutated kinases. 6j also demonstrated reasonable pharmacokinetic properties and a promising oral therapeutic effect in a bleomycin-induced mouse pulmonary fibrosis model.

Full text of DOI:10.1021/acs.jmedchem.6b00140

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Brief Article
Structure-Based Design of Tetrahydroisoquinoline-7-Carboxamides
as Selective Discoidin Domain Receptor 1 (DDR1) Inhibitors
Zhen Wang, Huan Bian, Sergio G. Bartual, Wenting Du, Jinfeng Luo, Hu Zhao,
Shasha Zhang, Cheng Mo, Yang Zhou, Yong Xu, Zheng-Chao Tu, Xiaomei Ren,
Xiaoyun Lu, Rolf A Brekken, Libo Yao, Alex N. Bullock, Jin Su, and Ke Ding
J. Med. Chem., Just Accepted Manuscript • DOI: 10.1021/acs.jmedchem.6b00140 • Publication Date (Web): 24 May 2016
Downloaded from http://pubs.acs.org on May 25, 2016
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Page 1 of 7
Journal of Medicinal Chemistry
DDR1 inhibition of the compounds was determined us-
was utilized as the potential hinge binding moiety. An N-
(3-((4-methylpiperazin-1-yl)methyl)-5-(trifluoromethyl)
phenyl)carboxamide group was also introduced based on
our previous investigation (Figure 2A).13
ing a well-established LANCE ULTRA kinase assay.25 The
potential target selectivity was additionally evaluated by
monitoring their inhibition against Abl. Compound 1 was
included as a positive control, which displayed similar
IC50 values to the previously reported data.13 It was shown
that 6a exhibited modest DDR1 inhibitory activity, with
an IC50 value of 442 nM, while its potency against Abl was
strikingly inferior (IC50 > 10.0 μM). These results are con-
sistent with our computational prediction.
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Scheme 1. Synthesis of Compound 6a and its Deriva-
B
C
tives.a
O
R1
R1
CF3
R1
NH2
N
H
R1
c
b
a
.Hcl
HN
F3
C
N
COOCH3
COOCH3
COOCH3
COOCH3
O
10
9
7
8
R1
R1
CF3
H
N
e
d
D
E
N
N
X
COOCH3
O
X
R2
Y
6a-6k
11
Y
aReagents and conditions: (a) Trifluoroacetic anhydride, 0
°C to rt, 58-63%; (b) (HCHO)n, conc. H2SO4, 0 °C to rt, 51-
89%; (c) (i) K2CO3, MeOH/H2O (2: 1), rt; (ii) HCl.MeOH,
MeOH, rt, 86∼95% (two steps); (d) 5-bromopyrimidine or 3-
bromopyridine or bromobenzene, Pd(dba)2, Ruphos, Cs2CO3,
toluene, 80 °C, 42-89%; (e) substituted aniline, t-BuOK, THF,
−20 °C to rt, 69−86%.
Figure 2. A) Design of new DDR1 inhibitor 6a. B) Molecular
docking of 6a (cyan) into the DDR1-ponatinib (yellow) co-
structure (PDB ID: 3ZOS). C) Co-crystal structure of 6c with
DDR1 (PDB ID: 5FDP). D) Molecular docking of 6b into
DDR1. E) Superposition of 6c with Abl (PDB ID: 3IK3).
Further computational investigation suggested that a
small hydrophobic recess formed by Val624, Ala653, and
Met699 was available in the ATP binding pocket of DDR1
(Figure 2B). A lipophilic substituent at R1 may occupy this
pocket to achieve improved potency. The (R)-methyl (6b)
and (R)-ethyl derivatives (6d) displayed a 20-fold and 12-
fold potency improvement, respectively. The (S)-methyl
compound (6c), in which the methyl moiety was oriented
away from the pocket, as confirmed by a 2.3 Å co-crystal
structure with DDR1 (Figure 2C), displayed a similar in-
hibitory potency to that of the R1 unsubstituted 6a. A
large group at the R1 position was predicted to be detri-
mental to the binding of DDR1. As expected, R1-(R)-
isopropyl (6f) caused a substantial potency loss for DDR1.
Additionally, compounds 6e and 6g, which featured (S)-
ethyl and (S)-isopropyl, respectively, displayed almost no
DDR1 inhibition. The X-ray crystal structure also con-
firmed the presence of a strong hydrogen bonding net-
work between the new inhibitor and DDR1 (Figure 2C).
The deletion of a hydrogen bond by eliminating the N
atoms in the pyrimidinyl group (6i) totally abolished
DDR1 inhibitory potency. Not surprisingly, when the py-
rimidinyl group was replaced by a pyridinyl moiety, the
resulting compound, 6h, exhibited an almost identical
IC50 value to that of 6a.
Our preliminary modeling suggested that the initial
lead, 6a, could fit nicely into the DDR1 binding pocket
and maintain the key interactions with DDR1. The pyrim-
idinyl moiety of 6a could form an essential hydrogen
bond with the NH of Met704 in the hinge region of DDR1.
Two additional hydrogen bonds have also formed be-
tween the linker amide and Glu672 in the C-helix and
Asp784 in the Asp-Phe-Gly (DFG) motif, respectively (Fig-
ure 2B). Encouragingly, compound 6a failed to dock into
the Abl binding pocket, suggesting its potential selectivity
among kinases. Compound 6a and its derivatives were
readily synthesized using Buchwald-Hartwig amination as
the key step (Scheme 1).24 Briefly, the substituted methyl
4-(2-(2,2,2-trifluoroacetamido)ethyl)benzoate (8) was
prepared by the trifluoroacetylation of methyl 4-(2-
aminoethyl)benzoate (7) and then underwent a classical
Pictet-Spengler reaction to yield the protected tetrahy-
droisoquinoline derivatives (9), which were deprotected
and reacted with a hydrochloric acid solution in MeOH to
form the key intermediate (10). Compound 10 was cou-
pled with 5-bromopyrimidine, 3-bromopyridine or bro-
mobenzene through a Buchwald-Hartwig amination reac-
tion to provide compound 11. The final products were
obtained by treating intermediate 11 with different ani-
lines under basic conditions.
It was also noteworthy that all the new DDR1 inhibitors
exhibited excellent DDR1 selectivity over the structurally
related Abl kinase (Table 1). To rationalize this target se-
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Page 2 of 7
lectivity, the inhibitor 6c was superimposed into the Abl
structure (PDB ID: 3IK3) (Figure 2E). It was shown that
the 1, 2, 3, 4-tetrahydroisoquinoline scaffold in 6c forced
the pyrimidinyl moiety to adopt a different dihedral angle
that prevented the formation of critical interactions with
Tyr253 and Phe382 in Abl. Moreover, the distance between
the N atom in the pyrimidinyl group and Met318 in Abl was
predicted to be 4.09 Å, which exceeds the limit to form a
potential hydrogen bond.
4-methyl-1H-
imidazol-1-yl
6k
113
N
N
(S)-Me
326±43
9.7±2.3
>10000
308±42
1
2
3
4
5
6
7
8
aDDR1 experiments were performed using the LANCE
ULTRA kinase assay, according to the manufacturer’s in-
structions. The data are the means from at least two inde-
pendent experiments. bAbl1 activity experiments were per-
formed using the FRET-based Z-Lyte assay, according to the
manufacturer’s instructions. The data are the means from at
least 3 independent experiments.
9
Table 1. The in vitro kinase inhibition of compounds 6a−6k
against DDR1a and Abl1b.
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Further structural optimization of the inhibitor 6b
yielded 6j as a promising candidate, with an IC50 value of
9.4 nM against DDR1 (Table 1). The compound also exhib-
ited reasonable pharmacokinetic (PK) properties, with an
oral bioavailability of 66.8% and a T1/2 value of 1.25 hours
at an oral dose of 20 mg/kg (Table 2) in rats. The PK pro-
file of compound 6j was also investigated in ICR mice,
which showed that the compound had a similar oral bioa-
vailability to that of rats. However, the area under con-
centration−time curve (AUC) value of the compound in
mice was obviously higher than that in rats, suggesting its
good absorption property in mice.
Kinase inhibition
Cpd
X
Y
R1
R2
(IC50, nM)
s
DDR1
Abl1
(4-
methylpiper-
azin-1-
yl)methyl
(4-
methylpiper-
azin-1-
yl)methyl
(4-
6a
N
N
H
442±69
24.3±4.1
309±44
>10000
Table 2. Pharmacokinetic profile of compound 6j in micea
and ratsb.
Mice
Rats
6b
6c
N
N
N
N
(R)-Me
(S)-Me
>10000
>10000
oral
4 mg/kg
iv
oral
iv
1 mg/kg 20 mg/kg 4 mg/kg
methylpiper-
azin-1-
AUC(0-∞)
788.3 ±
2554.1
1211.9
236.0 ± 53.1
(ng/mL.h)
41.5
yl)methyl
(4-
T1/2 (h)
1.1
0.5
0.2
1.3 ± 0
1 ± 0
1.5 ± 0.3
--
methylpiper-
azin-1-
6d
N
N
(R)-Et
36.4±5.7
>10000
Tmax (h)
--
2246.8
0.8
yl)methyl
Cmax
(ng/mL)
CLz
341.7 ±
115.5
473.3 ±
114.7
2193.9
(4-
methylpiper-
azin-1-
yl)methyl
(4-
methylpiper-
azin-1-
yl)methyl
(4-
methylpiper-
azin-1-
yl)methyl
(4-
methylpiper-
azin-1-
--
--
17.6 ± 4.4
6e
6f
N
N
N
N
N
N
N
C
(S)-Et
(R)-i-Pr
(S)-i-Pr
H
>2000
>1000
>3000
328±35
>10000
>10000
>10000
>10000
(L/h/kg)
BA (%)
41.6%
--
66.8%
--
aICR mice (male, 24 animals per group) weighing 18-30 g
were used for the study. bSD rats (male, 3 animals per
group) weighing 180−220 g were used for the study.
The DDR1 inhibition of 6j was further validated by de-
termining its binding affinity with the DDR1 protein
(conducted by DiscoveRx, San Diego, CA).26 It was shown
that 6j bound tightly to DDR1, with a binding constant
(Kd) value of 4.7 nM. The target specificity of 6j was also
investigated by conducting a kinase selectivity profiling
study against a panel of 468 kinases (including 403 non-
mutated kinases) at 1.0 μM, which is approximately 210-
fold above its Kd value against DDR1 using the DiscoveRx
screening platform. It was shown that 6j displayed excel-
lent target selectivity, with S(10) and S(1) scores of 0.022
and 0.012, respectively (Table S5).26 The potential “off-
target” kinases tested included cyclin-dependent kinase 11
(CDK 11), DDR2, Ephrin type-B receptor 8 (EPHB8), mus-
6g
6h
yl)methyl
(4-
methylpiper-
azin-1-
yl)methyl
6i
6j
C
C
H
>3000
>10000
>10000
4-methyl-1H-
imidazol-1-yl 9.4±1.7
N
N
(R)-Me
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Journal of Medicinal Chemistry
by Masson's trichrome, the lungs from BLM-challenged
cle-specific receptor tyrosine kinase (MUSK), nerve
1
2
3
4
5
6
7
8
growth factor receptor A (TrkA), TrkB, and TrkC. Howev-
er, the IC50 value of 6j against DDR2 was determined to be
188 nM in our LANCE ULTRA kinase assay, indicating
that 6j was 20-fold less potent against DDR2. Further de-
termination of the binding affinities (Kd values) revealed
that 6j exhibited an approximately 21-120-fold less poten-
cy against the majority of the other “off target” kinases,
with the exception of TrkB and TrkC, which displayed Kd
values of 22 and 18 nM, respectively (Figure 3). These re-
sults collectively supported the extraordinary target selec-
tivity of 6j against DDR1.
animals exhibited a reduction in alveolar spaces and were
stained blue by Masson's trichrome, demonstrating typi-
cal fibrotic features. The compound prevented these
BLM-induced pathological changes in a dose-dependent
manner (Figure 5A). These results agreed with the expres-
sion levels of fibrotic markers in lung tissue lysates, in-
cluding fibronectin and α-smooth muscle actin (SMA)
28
(Figure 5B). Further analyses also revealed that the ad-
9
ministration of compound 6j caused a dose-dependent
suppression in the content of hydroxyproline (Figure 5C),
a unique amino acid found in collagen.29 The above data
collectively indicate the promising therapeutic potential
of 6j against the BLM-induced pulmonary fibrosis.
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A
B
A
Figure 3. (A) KinomeScan kinase selectivity profiles for 6j.
Compound 6j was profiled at a concentration of 1.0 μM
against a diverse panel of 468 kinases by DiscoveRx. (B)
Binding constants (Kd values) of compound 6j against the top
hits. The data are the means from at least 3 independent
experiments.
B
C
Figure 5. Compound 6j prevents BLM-induced lung fibrosis.
Fourteen days after the onset of BLM injury, C57BL/6 mice
(
five animals each group) received an oral gavage of 6j twice
daily at the indicated dosages, and the lungs were collected
on day 28. The upper panels in (A) display the hematoxylin
and eosin (H&E) staining images of the dissected lungs. The
bottom panels in (A) represent Masson's trichrome staining.
The arrows indicate the fibrotic areas of the tissues. The
images in (B) show the results of the immunoblotting with
the indicated antibodies. The histogram in (C) shows the
determined hydroxyproline content. ***P<0.001.
Figure 4. Effects of DDR1 inhibition by 6j on signaling in
primary human lung fibroblasts. 6j inhibited DDR1-mediated
signaling in a concentration-dependent manner in primary
human lung fibroblasts (24-hour treatment). Lysates were
probed for the indicated targets by Western blot analysis.
CONCLUSION
Further investigation revealed that the activation of
DDR1 as well as its downstream signaling intermediate,
p38,27 were both dose-dependently suppressed by 6j in
primary human lung fibroblasts (Figure 4), suggesting the
efficacy of this compound against DDR1-induced signal-
ing. In light of the critical role of DDR1 in BLM-induced
pulmonary fibrosis,12 we treated mice with compound 6j
after the onset of a BLM challenge. Inhibitor 6j was orally
administered at 10 and 50 mg/kg twice daily (BID) for two
weeks based on its PK properties (Table 2). Unlike those
from the phosphate buffered saline (PBS)-treated mice,
which had large alveolar spaces and were weakly stained
In summary, a series of 1, 2, 3, 4-tetrahydroisoquinoline
derivatives were designed as novel highly selective DDR1
inhibitors. Compound 6j strongly suppressed DDR1, with
a single digital nM IC50 value, but it is significantly less
potent in a panel of 400 non-mutated kinases. Thus, to
the best of our knowledge, this compound represents one
of the most selective DDR1 inhibitors to date. The com-
pound also demonstrated reasonable PK properties and a
promising oral therapeutic effect in a BLM-induced
mouse pulmonary fibrosis model. Its strong DDR1 inhibi-
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Page 4 of 7
tyrosine; Phe, phenylalanine; Met, methionine; Glu, glutamic
tory potency and extraordinary target specificity make
compound 6j not only a promising lead compound for
new drug discovery but also a valuable research probe for
further biological investigation of its target.
1
2
3
4
5
6
7
8
acid; Asp, aspartic acid; DFG, Asp-Phe-Gly; MeOH, metha-
nol; PDB, Protein Data Bank; rt, room temperature;
Pd(dba)2, bis(dibenzylideneacetone)palladium; Ruphos, 2-
dicyclohexyl phosphino-2',6'-diisopropoxy-1,1'-biphenyl; t-
BuOK, potassium tert-butanolate; THF, tetrahydrofuran; Val,
valine; Ala, alanine; Cpds, compounds; AUC, area under con-
centration−time curve; T1/2, half-life period; ICR, Institute of
EXPERIMENTAL SECTION
General Chemistry. Reagents and solvents were obtained
from commercial suppliers and used without further purifi-
cation. Flash chromatography was performed using silica gel
Cancer Research; SD, Sprague Dawley; Tmax, peak time; Cmax
,
13
peak concentration; CL, clearance; BA, bioavailability; iv,
intravenous; CDK11, cyclin-dependent kinase 11, EPHB8,
ephrin type-B receptor 8; MUSK, muscle-specific receptor
(200−300 mesh). 1H and C NMR spectra were recorded on a
9
Bruker AV-400 spectrometer at 400 MHz and Bruker AV-500
spectrometer at 125 MHz. The low or high resolution of ESI-
MS was recorded on an Agilent 1200 HPLC-MSD mass spec-
trometer or Applied Biosystems Q-STAR Elite ESI-LC-
MS/MS mass spectrometer, respectively. The purity of com-
pounds was determined to be over 95% (>95%) by reverse-
phase high performance liquid chromatography (HPLC)
analysis. HPLC instrument: DIONEX SUMMIT HPLC (col-
umn, Diamonsil C18, 5.0 μm, 4.6 mm × 250 mm (Dikma
Technologies); detector, PDA-100 photodiode array; injector,
ASI-100 autoinjector; pump: p-680A). Elution: 85% MeOH in
water; flow rate, 1.0 mL/min.
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tyrosine kinase; TrkA, nerve growth factor
receptor A;
PHLF, primary human lung fibroblast; BLM, bleomycin; BID,
twice daily; PK, pharmacokinetic; PBS, phosphate buffered
saline; SMA, αꢀsmooth muscle actin; H&E, hematoxylin and
eosin.
ANCILLARY INFORMATION
Supporting Information
The supporting information includes synthetic procedures
and compound characterization, procedures and results for
in vitro kinase assay, KINOMEscanTM, protein expression and
purification, crystallization and structure determination,
computational study, western blot analysis, animal experi-
AUTHOR INFORMATION
Corresponding Authors
1
ments, antitumor activity of compound 6j. The H and 13C
E-mail: ding_ke@gibh.ac.cn; Tel: +86-20-32015276; Fax: +86-
20-32015299 (K. Ding)
NMR spectra of compounds 6a-6k, and molecular formula
strings. This material is available free of charge via the inter-
net at http://pubs.acs.org.
Atomic Coordinates and experimental data for the co-crystal
structure of 6c with DDR1 (PDB ID: 5FDP) will be released
upon article publication.
E-mail: sujin923@fmmu.edu.cn (J. Su)
E-mail: alex.bullock@sgc.ox.ac.uk (A. N. Bullock)
Author Contributions
‡
Z. Wang, H. Bian, and S. G. Bartual contributed equally to
REFERENCES
this work.
(1) Shrivastava, A.; Radziejewski, C.; Campbell, E.; Kovac,
L.; McGlynn, M.; Ryan, T. E.; Davis, S.; Goldfarb, M. P.;
Glass, D. J.; Lemke, G.; Yancopoulos, G. D. An orphan re-
ceptor tyrosine kinase family whose members serve as
nonintegrin collagen receptors. Mol. Cell 1997, 1, 25-34.
Notes
The authors declare no competing financial interests.
ACKNOWLEDGMENT
(2) Vogel, W.; Gish, G. D.; Alves, F.; Pawson, T. The dis-
coidin domain receptor tyrosine kinases are activated by
collagen. Mol. Cell 1997, 1, 13-23.
The authors appreciate the financial support from the Na-
tional Natural Science Foundation of China (81425021 and
21572230) and the Natural Science Foundation of Guangdong
Province (2015A03031201). We also thank Diamond Light
Source for beam time (proposal mx8421), as well as the staff
of beamline I04 for their assistance with crystal testing and
data collection. The SGC is a registered charity (number
1097737) that receives funds from AbbVie, Bayer Pharma AG,
Boehringer Ingelheim, the Canada Foundation for Innova-
tion, the Eshelman Institute for Innovation, Genome Canada,
Innovative Medicines Initiative (EU/EFPIA) [ULTRA-DD
grant no. 115766], Janssen, Merck & Co., Novartis Pharma AG,
Ontario Minis-try of Economic Development and Innovation,
Pfizer, São Paulo Research Foundation-FAPESP, Takeda, and
Wellcome Trust [092809/Z/10/Z].
(3) Xu, H.; Raynal, N.; Stathopoulos, S.; Myllyharju, J.;
Farndale, R. W.; Leitinger, B. Collagen binding specificity
of the discoidin domain receptors: binding sites on colla-
gens II and III and molecular determinants for collagen IV
recognition by DDR1. Matrix Biol. 2011, 30, 16-26.
(4) Vogel, W. F.; Abdulhussein, R.; Ford, C. E. Sensing ex-
tracellular matrix: an update on discoidin domain receptor
function. Cell Signal 2006, 18, 1108-1116.
(5) Valiathan, R. R.; Marco, M.; Leitinger, B.; Kleer, C. G.;
Fridman, R. Discoidin domain receptor tyrosine kinases:
new players in cancer progression. Cancer Metast. Rev.
2012, 31, 295-321.
ABBREVIATIONS
(6) Leitinger, B. Discoidin domain receptor functions in
physiological and pathological conditions. Cell Mol. Biol.
2014, 310, 39-87.
DDR, discoidin domain receptor; IC50, the half maximal
(50%) inhibitory concentration of a substance; RTKs, recep-
tor tyrosine kinases; p38 MAPK, P38 mitogen-activated pro-
tein kinase; Abl, abelson; ATP, adenosine triphosphate; Tyr,
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(20) Cowan-Jacob, S. W.; Fendrich, G.; Floersheimer, A.;
(7) Iwai, L. K.; Luczynski, M. T.; Huang, P. H. Discoidin
1
2
3
4
5
6
7
8
domain receptors: a proteomic portrait. Cell Mol. Life Sci.
2014, 71, 3269-3279.
Furet, P.; Liebetanz, J.; Rummel, G.; Rheinberger, P.; Cen-
teleghe, M.; Fabbro, D.; Manley, P. W. Structural biology
contributions to the discovery of drugs to treat chronic
myelogenous leukaemia. Acta Crystallogr. 2007, 63, 80-93.
(8) Borza, C. M.; Pozzi, A. Discoidin domain receptors in
disease. Matrix Biol. 2014, 34, 185-192.
(21) Weisberg, E.; Manley, P. W.; Breitenstein, W.; Bruggen,
J.; Cowan-Jacob, S. W.; Ray, A.; Huntly, B.; Fabbro, D.;
Fendrich, G.; Hall-Meyers, E.; Kung, A. L.; Mestan, J.; Da-
ley, G. Q.; Callahan, L.; Catley, L.; Cavazza, C.; Azam, M.;
Neuberg, D.; Wright, R. D.; Gilliland, D. G.; Griffin, J. D.
Characterization of AMN107, a selective inhibitor of native
and mutant Bcr-Abl. Cancer cell 2005, 7, 129-141.
(9) Kothiwale, S.; Borza, C. M.; Lowe, E. W., Jr.; Pozzi, A.;
Meiler, J. Discoidin domain receptor 1 (DDR1) kinase as
target for structure-based drug discovery. Drug Discov.
Today 2015, 20, 255-261.
9
(10) Ju, G. X.; Hu, Y. B.; Du, M. R.; Jiang, J. L. Discoidin
domain receptors (DDRs): potential implications in ather-
osclerosis. Eur. J. Pharmacol. 2015, 751, 28-33.
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54
55
56
57
58
59
60
(22) O'Hare, T.; Shakespeare, W. C.; Zhu, X.; Eide, C. A.;
Rivera, V. M.; Wang, F.; Adrian, L. T.; Zhou, T.; Huang, W.
S.; Xu, Q.; Metcalf, C. A., 3rd; Tyner, J. W.; Loriaux, M. M.;
Corbin, A. S.; Wardwell, S.; Ning, Y.; Keats, J. A.; Wang, Y.;
Sundaramoorthi, R.; Thomas, M.; Zhou, D.; Snodgrass, J.;
Commodore, L.; Sawyer, T. K.; Dalgarno, D. C.; Deininger,
M. W.; Druker, B. J.; Clackson, T. AP24534, a pan-BCR-ABL
inhibitor for chronic myeloid leukemia, potently inhibits
the T315I mutant and overcomes mutation-based re-
sistance. Cancer cell 2009, 16, 401-412.
(11) Li, Y.; Lu, X.; Ren, X.; Ding, K. Small molecule dis-
coidin domain receptor kinase inhibitors and potential
medical applications. J. Med. Chem. 2015, 58, 3287-3301.
(12) Avivi-Green, C.; Singal, M.; Vogel, W. F. Discoidin do-
main receptor 1-deficient mice are resistant to bleomycin-
induced lung fibrosis. AM. J. Resp. Crit. Care Med. 2006,
174, 420-427.
(13) Gao, M.; Duan, L.; Luo, J.; Zhang, L.; Lu, X.; Zhang, Y.;
Zhang, Z.; Tu, Z.; Xu, Y.; Ren, X.; Ding, K. Discovery and
optimization of 3-(2-(Pyrazolo[1,5-a]pyrimidin-6-yl)ethyn
yl)benzamides as novel selective and orally bioavailable
discoidin domain receptor 1 (DDR1) inhibitors. J. Med.
Chem. 2013, 56, 3281-3295.
(23) Canning, P.; Tan, L.; Chu, K.; Lee, S. W.; Gray, N. S.;
Bullock, A. N. Structural mechanisms determining inhibi-
tion of the collagen receptor DDR1 by selective and multi-
targeted type II kinase inhibitors. J. Mol. Biol. 2014, 426,
2457-2470.
(14) Kim, H. G.; Tan, L.; Weisberg, E. L.; Liu, F.; Canning,
P.; Choi, H. G.; Ezell, S. A.; Wu, H.; Zhao, Z.; Wang, J.;
Mandinova, A.; Griffin, J. D.; Bullock, A. N.; Liu, Q.; Lee, S.
W.; Gray, N. S. Discovery of a potent and selective DDR1
receptor tyrosine kinase inhibitor. ACS Chem. Biol. 2013, 8,
2145-2150.
(24) Surry, D. S.; Buchwald, S. L. Dialkylbiaryl phosphines
in Pd-catalyzed amination: a user's guide. Chem. Sci. 2011,
2, 27-50.
(25) The assay was conducted by following the protocol
from
the
agent
manufacturer
(15) Elkamhawy, A.; Park, J. E.; Cho, N. C.; Sim, T.; Pae, A.
N.; Roh, E. J. Discovery of a broad spectrum antiprolifera-
tive agent with selectivity for DDR1 kinase: cell line-based
assay, kinase panel, molecular docking, and toxicity stud-
ies. J. Enzym. Inhib. Med. Chem. 2016, 31, 158-166.
(https://perkinelmerreagent s.onconfluence.com)
(26) Fabian, M. A.; Biggs, W. H., 3rd; Treiber, D. K.; Atter-
idge, C. E.; Azimioara, M. D.; Benedetti, M. G.; Carter, T.
A.; Ciceri, P.; Edeen, P. T.; Floyd, M.; Ford, J. M.; Galvin,
M.; Gerlach, J. L.; Grotzfeld, R. M.; Herrgard, S.; Insko, D.
E.; Insko, M. A.; Lai, A. G.; Lelias, J. M.; Mehta, S. A.; Mila-
nov, Z. V.; Velasco, A. M.; Wodicka, L. M.; Patel, H. K.; Zar-
rinkar, P. P.; Lockhart, D. J. A small molecule-kinase inter-
action map for clinical kinase inhibitors. Nat. Biotechnol.
2005, 23, 329-336.
(16) Richters, A.; Nguyen, H. D.; Phan, T.; Simard, J. R.;
Grutter, C.; Engel, J.; Rauh, D. Identification of type II and
III DDR2 inhibitors. J. Med. Chem. 2014, 57, 4252-4262.
(17) Terai, H.; Tan, L.; Beauchamp, E. M.; Hatcher, J. M.;
Liu, Q.; Meyerson, M.; Gray, N. S.; Hammerman, P. S.
Characterization of DDR2 inhibitors for the treatment of
DDR2 mutated non-small cell lung cancer. ACS Chem.
Biol. 2015, 10, 2687-2696.
(27) Matsuyama, W.; Wang, L.; Farrar, W. L.; Faure, M.;
Yoshimura, T. Activation of discoidin domain receptor 1
isoform b with collagen up-regulates chemokine produc-
tion in human macrophages: role of p38 mitogen-activated
protein kinase and NF-kappa B. J. Immunol. 2004, 172,
2332-2340.
(18) Murray, C. W.; Berdini, V.; Buck, I. M.; Carr, M. E.;
Cleasby, A.; Coyle, J. E.; Curry, J. E.; Day, J. E.; Day, P. J.;
Hearn, K.; Iqbal, A.; Lee, L. Y.; Martins, V.; Mortenson, P.
N.; Munck, J. M.; Page, L. W.; Patel, S.; Roomans, S.;
Smith, K.; Tamanini, E.; Saxty, G. Fragment-based discov-
ery of potent and selective DDR1/2 inhibitors. ACS Med.
Chem. Lett. 2015, 6, 798-803.
(28) Nagase T.; Uozumi N.; Ishii S.; Kita Y.; Yamamoto H.;
Ohga E.; Ouchi Y.; Shimizu T. A pivotal role of cytosolic
phospholipase A (2) in bleomycin-induced pulmonary
fibrosis. Nat Med. 2002, 8, 480-484.
(19) Day, E.; Waters, B.; Spiegel, K.; Alnadaf, T.; Manley, P.
W.; Buchdunger, E.; Walker, C.; Jarai, G. Inhibition of col-
lagen-induced discoidin domain receptor 1 and 2 activa-
tion by imatinib, nilotinib and dasatinib. Eur. J. Pharma-
col. 2008, 599, 44-53.
(29) Colgrave, M. L.; Allingham, P. G.; Tyrrell, K.; Jones, A.
Multiple reaction monitoring for the accurate quantifica-
tion of amino acids: using hydroxyproline to estimate col-
lagen content. Methods Mol. Biol. 2012, 828, 291-303.
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