Discovery and Pharmacokinetic and Pharmacological Properties of the Potent and Selective MET Kinase Inhibitor 1-{6-[6-(4-Fluorophenyl)-[1,2,4]triazolo[4,3-b]pyridazin-3-ylsulfanyl]benzothiazol-2-yl}-3-(2-morpholin-4-ylethyl)urea (SAR125844)

Article abstract of DOI:10.1021/acs.jmedchem.6b00280

The HGF/MET pathway is frequently activated in a variety of cancer types. Several selective small molecule inhibitors of the MET kinase are currently in clinical evaluation, in particular for NSCLC, liver, and gastric cancer patients. We report herein the

Full text of DOI:10.1021/acs.jmedchem.6b00280

Subscriber access provided by - Access paid by the | UCSB Libraries
Drug Annotation
Discovery, pharmacokinetic and pharmacological properties
of the potent and selective MET kinase inhibitor, 1-{6-[6-(4-
Fluoro-phenyl)-[1,2,4]triazolo[4,3-b]pyridazin-3-ylsulfanyl]-
benzothiazol-2-yl}-3-(2-morpholin-4-yl-ethyl)-urea (SAR125844)
Antonio Ugolini, Mireille Kenigsberg, Alexey Rak, François Vallée, Jacques Houtmann,
Maryse Lowinski, Cécile Capdevila, Jean Khider, Eva Albert, Nathalie Martinet, Conception
Nemecek, Sandrine Grapinet, Eric Bacqué, Manfred Roesner, Christine Delaisi, Loreley Calvet,
Fabrice Bonche, Dorothée Semiond, Coumaran Egile, Hélène Goulaouic, and Laurent Schio
J. Med. Chem., Just Accepted Manuscript • DOI: 10.1021/acs.jmedchem.6b00280 • Publication Date (Web): 29 Jun 2016
Downloaded from http://pubs.acs.org on June 30, 2016
Just Accepted
“Just Accepted” manuscripts have been peer-reviewed and accepted for publication. They are posted
online prior to technical editing, formatting for publication and author proofing. The American Chemical
Society provides “Just Accepted” as a free service to the research community to expedite the
dissemination of scientific material as soon as possible after acceptance. “Just Accepted” manuscripts
appear in full in PDF format accompanied by an HTML abstract. “Just Accepted” manuscripts have been
fully peer reviewed, but should not be considered the official version of record. They are accessible to all
readers and citable by the Digital Object Identifier (DOI®). “Just Accepted” is an optional service offered
to authors. Therefore, the “Just Accepted” Web site may not include all articles that will be published
in the journal. After a manuscript is technically edited and formatted, it will be removed from the “Just
Accepted” Web site and published as an ASAP article. Note that technical editing may introduce minor
changes to the manuscript text and/or graphics which could affect content, and all legal disclaimers
and ethical guidelines that apply to the journal pertain. ACS cannot be held responsible for errors
or consequences arising from the use of information contained in these “Just Accepted” manuscripts.
Journal of Medicinal Chemistry is published by the American Chemical Society. 1155
Sixteenth Street N.W., Washington, DC 20036
Published by American Chemical Society. Copyright © American Chemical Society.
However, no copyright claim is made to original U.S. Government works, or works
produced by employees of any Commonwealth realm Crown government in the course
of their duties.

Page 1 of 22
Journal of Medicinal Chemistry
1
2
3
4
5
6
7
8
9
Discovery, pharmacokinetic and pharmacological
properties of the potent and selective MET kinase
inhibitor, 1ꢀ{6ꢀ[6ꢀ(4ꢀFluoroꢀphenyl)ꢀ[1,2,4]triazolo[4,3ꢀ
b]pyridazinꢀ3ꢀylsulfanyl]ꢀbenzothiazolꢀ2ꢀyl}ꢀ3ꢀ(2ꢀ
morpholinꢀ4ꢀylꢀethyl)ꢀurea (SAR125844)
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
Antonio Ugolini,† Mireille Kenigsberg, ¶ Alexey Rak, ‡ Francois Vallée,‡ Jacques Houtmann,‡ Maryse
Lowinski,‡ Cécile Capdevila, µ Jean Khider,† Eva Albert,† Nathalie Martinet,† Conception Nemecek,†
Sandrine Grapinet,† Eric Bacqué,†,φ Manfred Roesner, ƪ Christine Delaisi,¶ Loreley Calvet, ¶ Fabrice
Bonche, † Dorothée Semiond, †,ƽ Coumaran Egile,¶,§ Hélène Goulaouic, ¶ Laurent Schio†,*
†Medicinal Chemistry, ¶ Oncology Drug Discovery, ‡ Structure Design & Informatics, µ Protein Science
& Technology, Sanofi, 13 quai Jules Guesde, 94403 VitryꢀsurꢀSeine, France. †Disposition Safety
Animal Research, 1 avenue Pierre Brossolette, 91385 ChillyꢀMazarin, France. ƪ Industriepark Hoechst,
65926 Frankfurt am Main, Germany.
Current address: §AstraZeneca, Oncology Innovative Medicines, Personalized Healthcare and
Biomarkers, United Kingdom; φ Infectious Diseases Unit, 1541 Avenue Marcel Merieux, 69280, Marcy
L’Etoile, France; ƽ 500 Kendall St, 2142 Cambridge, MA, United States.
ABSTRACT: The HGF/MET pathway is frequently activated in a variety of cancer types. Several
selective small molecule inhibitors of the MET kinase are currently in clinical evaluation, in particular
ACS Paragon Plus Environment
1

Journal of Medicinal Chemistry
Page 2 of 22
for NSCLC, liver and gastric cancer patients. We report herein the discovery of a series of
triazolopyridazines that are selective inhibitors of wildꢀtype (WT) MET kinase and several clinically
relevant mutants. We provide insight into their mode of binding and report unprecedented crystal
structures of the Y1230H variant. A multiꢀparametric chemical optimization approach allowed the
identification of compound 12 (SAR125844) as a development candidate. In this chemical series,
absence of CYP3A4 inhibition was obtained at the expense of satisfactory oral absorption. Compound
12, a promising parenteral agent for the treatment of METꢀdependent cancers, promoted sustained target
engagement at tolerated doses in a human xenograft tumor model. Preclinical pharmacokinetics
conducted in several species were predictive for the observed pharmacokinetic behavior of 12 in
patients.
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
KEYWORDS: cꢀMet, MET kinase inhibitor, MET amplification, SAR125844, triazolopyridazine,
multiꢀparametric optimization, Y1230H Xꢀray structure.
The MET receptor tyrosine kinase is activated by binding of its ligand, Hepatocyte Growth Factor
(HGF), resulting in receptor dimerization, autoꢀphosphorylation of Tyr1234/1235, activation of the
kinase domain and of the downstream multiple signaling pathways, including RAS/MAPK and
PI3K/AKT, which in turn promote cell proliferation, survival, motility, invasion, angiogenesis and
morphogenesis.1,2 The HGF/MET pathway plays an essential physiological role during embryogenesis3,
and a more restricted role in tissue regeneration and damage repair in healthy adults.4,5 Dysregulation of
MET has been reported in several tumor types, either in the form of overexpression of the MET
receptor, or as mutations6, 7 and amplification8 of the MET gene. Importantly, MET gene amplification
was reported as an acquired resistance mechanism to EGFR tyrosine kinase inhibitor therapies in 10ꢀ
20% of NonꢀSmall Cell Lung Cancer (NSCLC) patients9,10,11 and to antiꢀEGFR monoclonal antibody
treatment in Colorectal Cancer (CRC) patients.12 More recently, HGF expression in the tumor stroma
was reported as a resistance mechanism to BRAF inhibitors.13
ACS Paragon Plus Environment
2

Page 3 of 22
Journal of Medicinal Chemistry
Several orally bioavailable MET tyrosine kinase inhibitors, including Capmatinib (INC280),14
Tepotinib (EMD1214063),15 Savolitinib (Volitinib/AZD6094/HMPLꢀ504),16 (R)ꢀ6ꢀ(1ꢀ(8ꢀFluoroꢀ6ꢀ(1ꢀ
methylꢀ1Hꢀpyrazolꢀ4ꢀyl)ꢀ[1,2,4]triazolo[4,3ꢀa]pyridinꢀ3ꢀyl)ethyl)ꢀ3ꢀ(2ꢀmethoxyethoxy)ꢀ1,6ꢀ
naphthyridinꢀ5(6H)ꢀone (AMGꢀ337)17 as well as the MET/ALK/ROS inhibitor Crizotinib,18 are in Phase
II clinical trials in papillary renal cell carcinoma, NSCLC, gastroꢀesophageal and hepatocellular cancer.
We report here the discovery, pharmacological characterization, preclinical and clinical
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
pharmacokinetics
of
1ꢀ{6ꢀ[6ꢀ(4ꢀFluoroꢀphenyl)ꢀ[1,2,4]triazolo[4,3ꢀb]pyridazinꢀ3ꢀylsulfanyl]ꢀ
potent
benzothiazolꢀ2ꢀyl}ꢀ3ꢀ(2ꢀmorpholinꢀ4ꢀylꢀethyl)ꢀurea, compound 12 (SAR125844),
a
intravenously active and highly selective MET kinase inhibitor; comparable to the aforementioned MET
specific clinical candidates as assessed through biochemical and cellular profiling.19, 20
The original hit identification approach was based on a mediumꢀthroughput screening of an inꢀhouse
biased kinase inhibitor library tested in a biochemical assay using nonꢀphosphorylated MET kinase
domain in order to identify all classes (DFG in (AspꢀPheꢀGly), DFG out, αꢀHelix out etc.) of potential
inhibitors.21 Subsequent biochemical assays were performed with phosphorylated MET kinase domain
during the optimization phase.
A set of benzimidazole sulfonate derivatives with nanomolar MET inhibition was identified. In
particular, the initial hit 1 (Figure 1) exhibited an IC50 of 5 nM against MET kinase but also displayed a
strong undesirable affinity for Cyclin Dependent Kinase CDK9 (IC50= 6 nM), an isoform involved in
gene transcription processes. Modification of the sulfonyl aryl moiety with more bulky substituents (e.g.
chlorine vs fluorine atoms in analog 2) was well tolerated in the MET protein (2: MET IC50 = 23 nM)
and resulted in prohibitive steric clashes and reduced binding to CDK9 (2: IC50= 1200 nM). However,
compound 2 also displayed potent tubulin affinity (IC50 = 2.7 µM, and microtubule poisons are active in
the 0.5ꢀ3 µM subꢀstoichiometric concentration range in a microtubule assembly assay performed with 6ꢀ
8 µM tubulin) leading to offꢀtarget cytotoxicity in nonꢀMETꢀdependent tumor cell lines.
ACS Paragon Plus Environment
3

Journal of Medicinal Chemistry
Page 4 of 22
Revisiting the internal compound collection retrieved a set of about 2000 benzimidazole derivatives
from which a potent singleton originating from a former anthelmintic program was identified by testing
on the phosphorylated MET kinase domain. Triazolopyridazine 3 was active (IC50 = 10 nM) in the MET
biochemical assay, the ELISA PꢀMET cellꢀbased assay as well as the proliferation assay using the
MKNꢀ45 METꢀamplified gastric tumor cell line. Compound 3 was inactive on CDK9 (IC50 > 10 µM)
but still displayed affinity toward tubulin (IC50 = 2 µM).
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
O
O
S
S
N
N
O
O
O
S
N
F
S
Cl
S
N
N
N
O
O
N
H
N
H
N
N
O
O
N
O
N
H
N
N
N
N
H
N
H
N
H
O
O
O
O
Cl
F
N
H
N
H
1
2
4
3
F
F
Figure 1. Hit and lead finding steps
To unravel the molecular interactions responsible for the kinase activity observed we initiated structural
characterization of compound 3 bound to the WT MET kinase domain. Coꢀcrystals were grown, Xꢀray
diffraction data collected (1.6Å resolution at the ESRF synchrotron ID 14ꢀ2 beamline in Grenoble,
France) and the 3D structure of the complex determined. Structure analysis revealed that compound 3
binds to the kinase ATP pocket, forming two Hꢀbonds with the hinge region (Met1160, Figure 2A). In
addition, one of the nitrogen atoms of the triazolopyridazine moiety interacts with the mainꢀchain
nitrogen atom of Asp1222. The triazolopyridazine ring makes a πꢀπ interaction with Tyr1230 of the
activation loop positioned in a nonꢀactive conformation. This binding mode is a structural feature of the
soꢀcalled type I inhibitors, as exemplified by Crizotinib and other triazolopyridazine / triazolopyridineꢀ
bearing MET inhibitors.22
Based on the coꢀcrystal structure of 3 bound to MET kinase, chemical modifications were designed and
performed to simultaneously improve (i) the potency against WT MET and MET mutant proteins, (ii)
the selectivity (among kinases and vs tubulin) and (iii) the aqueous thermodynamic solubility. The latter
physicochemical property was assessed under two conditions, in pure buffer (intrinsic solubility) and
ACS Paragon Plus Environment
4

Page 5 of 22
Journal of Medicinal Chemistry
with a coꢀsolvent (labrasol) as a preliminary formulation. In the course of the optimization program the
benzothiazole subꢀseries (e.g. compound 4, Figure 1 and Table 1) stood out as more attractive than the
corresponding benzimidazoles, since it was not binding to tubulin (IC50 > 25µM) and had no Ames alert
(compound 3 was found Ames positive). However, switching to the benzothiazole series was at risk
from a druggability standpoint as compound 4 was more lipophilic compared to 3 (logD = 4.6 versus
3.8) and exhibited decreased aqueous solubility in the conditions tested.
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
A.
B.
Figure 2. Xꢀray structures of WT MET with 3 (A) and METꢀY1230H with 14 (B)
Various chemical moieties (carbamates, amines, amides and ureas) were introduced in position 2 of
the benzothiazole scaffold allowing at least a double interaction with the hinge region of the kinase (R1
group in Table 1). Most of the modifications depicted in Table 1 were well tolerated leading to potent
kinase inhibitions in the biochemical assay, which consistently translated into significant PꢀMET
inhibition in the cellꢀbased assay, as well as antiꢀproliferative activity on the METꢀamplified MKNꢀ45
cell line (IC50 values of 10ꢀ100 nM). Although these compounds were found to be significantly active
against the M1250T mutant, they revealed to be less potent when considering L1195V and Y1230H
mutants (see compounds 4, 5, 6 &7 vs 8 and 9 in Table 1). Interestingly, the most catalytically active
mutants were also the most sensitive ones (data not shown). In addition subꢀmicromolar inhibition of the
Y1230H mutant was achieved only with compounds that were active at least in the nanomolar range
against WT MET protein (compounds 9, 12 & 13). From a structureꢀpotency relationship point of view,
introduction of a third interaction between the additional nitrogen atom brought by the urea group and
ACS Paragon Plus Environment
5

Journal of Medicinal Chemistry
Page 6 of 22
the hinge does not explain the observed gain of activity for ureas 12 and 13 (compared to 4) since
cyclopropyl amide 9 is equipotent to ureas 12 and 13. To better understand the thermodynamics of
binding in the selected series, we performed Isothermal Titration Calorimetry (ITC) measurements23
with a series of analogs including the most active compound 12. In general, a good correlation was
1
2
3
4
5
6
7
8
9
observed between the biochemical IC50 and the Kd obtained by calorimetry (e.g. Kd = 0.9 nM vs IC50 =
4nM for 12). The free energy of binding of compound 12 to the WT protein was measured
experimentally and found to be totally driven by the enthalpy contribution (ꢁG=ꢁH= ꢀ12.6 Kcal/mol), a
trend shared by the different triazolopyridazine compounds tested and in contrast to the ITC profile of
Crizotinib (Figure 3).
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
H
H
H
H
N
O
N
N
N
O
N
O
N
O
N
N
N
N
N
N
O
O
NH
O
NH
NH
NH
S
NH
O
S
S
S
N
N
N
S
S
H2N
S
S
N
N
N
N
N
N
N
N
O
N
N
N
N
N
N
N
N
Cl
F
O
F
Cl
Crizotinib
F
15
18
17
12 (SAR125844)
Figure 3: Structures of key compounds with biochemical and ITC data.
This raised the hypothesis that potency was directly dependent on the number of hydrogen bonds, Van
der Waals interactions and hydrophobic contacts24 the inhibitor could establish with the protein. To gain
insight on improving the affinity for MET mutants, coꢀcrystallization attempts with compound 12 were
performed on Y1230H, L1195V and M1250T mutant proteins, and with MET WT as well. Surprisingly,
ACS Paragon Plus Environment
6

Page 7 of 22
Journal of Medicinal Chemistry
whereas the WT protein did not yield any crystals, 3D structures were readily obtained with the
different MET mutants. Structural analysis of different complexes with 12 revealed 4 conserved major
hydrogen bonds, three developed with Met1160 and Lys1161 of the hinge and one with Asp1222 of the
DFG motif.
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
Figure 4: Crystal structure of 12 in the Y1230H mutant showing the tight interaction with the hinge and the DFG
Asp1222.
On the other hand, the activation loop segment containing residue 1230 was not visible in any
structure with 12 (supplementary information). Similarly to the 3D structure of 3 in WT MET protein,
the mutated proteins were locked in a DFG in, αꢀC helix out conformation with a disrupted salt bridge
between catalytic Lys1110 and αꢀC Helix Glu112725. Nonꢀvisibility of the H1230 residue bearing
segment in the MET Y1230H coꢀcrystal structure raised the question whether reduced affinity of
compound 12 (and other analogs) for this mutant might be linked to a subꢀoptimal πꢀπ interaction of the
triazolopyridazine core with the imidazole ring of H1230. This hypothesis was confirmed when
compound 14 (analog of 12) was successfully coꢀcrystallized in the MET Y1230H mutant, the structure
of the complex showing a clear positioning of the histidine residue in the electron density maps (Figure
2B). In this crystal structure, the triazolopyridazine plane is twisted outward by ~20° with respect to
other analogs, thus placing the thiophene ring roughly parallel to H1230. Also observed is a slight shift
of H1230 (~1Å) towards the solvent compared to Y1230 as seen in other WT MET structures. High
potency of 14 against MET Y1230H mutant (IC50 = 23 nM) could then be explained by favorable
ACS Paragon Plus Environment
7

Journal of Medicinal Chemistry
Page 8 of 22
hydrophobic contacts between H1230 and the triazolopyridazineꢀthiophene segment (extended πꢀ
system). This hydrophobic interaction is likely enhanced by the planarity of the triazolopyridazineꢀ
thiophene segment whereas the skewed nature of the triazolopyridazineꢀpꢀfluorophenyl observed in 3
probably also precludes a productive contact in 12 (supplementary information).
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
Table 1. Biochemical and cellular activities on wildꢀtype MET, biochemical activity on MET mutants,
and aqueous solubility of benzothiazole derivatives 4-14
S
N
S
N
N
R1
N
N
F
Aqueous
Solubility
ELISA
P-MET
IC50
P-MET
WT IC50
(nM)a
MKN-45
Proliferation
IC50 (nM)c
L1195V
M1250T Y1230H
logD
Ref.g
(µM, pH7.4)f
+10% Labrasol
Compound
R1
IC50 (nM)d IC50 (nM)d IC50 (nM)d (pH7.4)e
(nM)b
4
5
6
NHCO2Me
NH2
16
10
15
22
43
15
91
81
42
>10000
9000
nd
>10000
>10000
>10000
>4.6
>4.6
>4.6
<1
nd
15
44
26
26
245
417
NHEt
H
N
N
>10000
<2.5
nd
*
7
20
6
13
2
45
12
7
835
318
455
93
13
128
nd
6
>10000
1172
883
4.1
4.3
<1
543
26
26
26
O
H
N
N
*
8
nd
<1
<2
8
96
64
O
O
H
N
*
9
3
4
61
4.7*
>4.6
4.1
O
H
N
10
11
12
13
14
*
4
13
25
5
22
95
7
>10000
6523
64
>10000
>10000
204
nd
O
H
N
O
N
N
*
45
4
245
500
1000
nd
26
26
26
O
O
O
H
N
H
N
*
4.2
<1
<1
<2
O
H
N
H
N
N
*
3
10
29
33
2.7
148
71
nd
864
4.15
3.9
O
S
N
S
N
H
N
N
H
N
N
N
O
2
nd
23
S
N
a IC50 measured using phosphorylated protein in a biochemical assay
ACS Paragon Plus Environment
8

Page 9 of 22
Journal of Medicinal Chemistry
b IC50 for cellꢀbased PꢀMET, using an ELISA assay and MKNꢀ45 cell line
c IC50 measured in proliferation assay using MKNꢀ45 cell line
1
2
3
4
d IC50 measured using MET mutants in a biochemical assay
see Supporting Information for details on assays a, b, c, d
5
6
7
8
e Measured experimentally by High Performance Liquid Chromatography (HPLC) (Xterra MS C18 column)
f Equilibrated solubility measured in phosphate buffer with and without 10% labrasol
g Compounds were prepared according to the methods in the reference cited
9
10
11
12
13
14
This observation prompted us to initiate a second round of chemical optimization with modifications
of the substituent linked to the triazolopyridazine core. The main purpose was to identify alternative
replacements of the thiophene and significantly improve the aqueous solubility. The reported activities
and drug like properties (solubility, permeability in Cacoꢀ2 cells, metabolic stability on human
microsomes, reversible and irreversible inhibition of CYP3A4) are shown in Table 2.
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
Compared to compound 15 (R2 = H), most of the substituents that were introduced led to potent analogs
against the WT MET protein; having variable activity against the MET Y1230H mutant. The lack of
permeability observed in the Cacoꢀ2 assay (Table 2) could be attributed to several unfavorable factors
like high molecular weight (in general around 550 g/mol), high number of Hꢀbond acceptors (at least 10
as in 15) and low intrinsic aqueous solubility; in spite of the balanced morpholino group basicity (pka =
6.5) suitable for passive diffusion.27 In contrast, 6ꢀOꢀalky analogs showed good to moderate Cacoꢀ2
permeability (compounds 18, 23 & 24). It remained unclear how to rationalize this observation since
these molecules exceed molecular weight (MW) of 500 g/mol, display 11 Hꢀbond acceptors and besides
13 and 26 the highest number of rotatable bonds (7) of this set of compounds.28 In line with the Cacoꢀ2
results, compounds 18 and 23 exhibited moderate bioavailability in mouse PK studies (17% and 12%,
respectively) as observed finally with many other extended rule of 5 (Ro5) compliant molecules.29
Unfortunately, these molecules were discarded due to their inhibitory activity on the CYP3A4 isoform
in a timeꢀdependent manner (mechanismꢀbased inhibition). Overall, taking into account the properties of
R2 derivatives reported in Table 2, compound 12 turned out to be the best compromise in terms of
potency and drugꢀlike properties in particular regarding CYP450 inhibition. In the end, attempts to
improve solubility in this chemical series failed as key atoms interacting with the MET proteins also
ACS Paragon Plus Environment
9

Journal of Medicinal Chemistry
Page 10 of 22
proved to be involved in strong packing forces in compound solid states. The single crystal Xꢀray
structure (Figure 5) of one polymorph of 12 attested to the dense packing of this solid state. The two flat
conjugated piꢀsystems disposed in a right angle configuration by the sulfur linker displayed stabilizing
Hꢀbond, lipophilic and πꢀstacking interactions (supplementary materials).
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
Figure 5: Unit cell and packing views in the Xꢀray single crystal diffraction structure of 12
In PK studies in mice, the oral bioavailability of 12 was low (~2%) in line with its moderate Cacoꢀ2
permeability. Hence, 12 was progressed toward in vivo preclinical studies as a parenteral agent for the
treatment of METꢀdependent cancer.
Table 2. Multiꢀparametric analysis of morpholino ethyl urea analogs26, 30
S
N
S
N
H
N
N
H
N
N
N
O
N
R2
O
aqueous
P-MET
WT
P-MET
Y1230H
IC50
Permea- % metabol.
Inhibition CYP3A4
solubility pH7.4 (µM)c
bility in
Caco-2
cellsd
in human
micro-
Compound
R2
H
of CYP3A4 inhibition Ref.h
IC50
Labrasol
(IC50, µM)f
(MBI)g
Aq. buffer
(nM) a
(nM)b
somee
15
29
4805
70
1070
0
11
34
-2.3
26
ACS Paragon Plus Environment
10

Page 11 of 22
Journal of Medicinal Chemistry
*
1
2
3
4
5
6
7
8
12
4
3
204
643
<2
5
500
460
9
0
23
51
>200
4
No*
26
26
F
OEt
16
17
-13.1
*
10
2
140
252
139
32
<2
<2
1170
500
ND
4
98
0
24
23
38
1.8
2.7
2.9
-8.2
-4.6
-9.1
26
F
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
O
*
*
*
18
19
30a
30a
NH
1
O
NH
20
1
217
89
ND
2
39
>40
-14.2
30c
NH
*
21
22
1
2
274
174
<2
<2
ND
ND
0
8
26
32
0.2
7.9
-3.8
30a
30b
OPh
-12.8
O
*
23
24
6
216
407
<2
<2
ND
ND
26
20
29
36
31
14
-5.5
30a
30a
O
*
*
15
-11.6
O
F
3
5
56
<2
<2
ND
ND
8
7
16
23
2
-12.2
-11.3
30b
25
26
O-nBu
140
8.7
a IC50 measured using preꢀphosphorylated wildꢀtype MET kinase domain in a biochemical assay
b IC50 measured using MET mutant Y1230H in a biochemical assay
c Equilibrated solubility measured in phosphate buffer with and without 10% labrasol
d TransꢀEpithelial Transport across Cacoꢀ2/ TCꢀ7 Monolayers (Papp = 10ꢀ7 cm.secꢀ1)
e Percentage of oxidative metabolism using Hepatic Microsomal Fraction (20 minutes incubation, 1 mg/mL microsomes,5µM substrate)
f IC50 measured using recombinant human CYP3A4 with 7ꢀbenzyloxyquinoline as substrate
g Negative Slope values in %/min reflect potency of the MechanismꢀBased Inhibition
h Compounds were prepared according to the methods in the references cited
See Supporting Information for details on assays a, b, d, e, f
* for IC50 higher than 100 ꢁM, no slopes can be determined and is defined as not quantifiable
ACS Paragon Plus Environment
11

Journal of Medicinal Chemistry
Page 12 of 22
The pharmacokinetic and pharmacodynamic properties of compound 12 were first documented in
1
2
3
4
5
6
7
8
9
female SCID mice bearing xenograft tumors based on the METꢀamplified Hs 746T human gastric tumor
cell line. To this end, a single intravenous administration at 20 mg/kg was performed. Compound 12
concentration was measured in both plasma and tumor tissue. The plasma exposure of 12 (Area Under
the Curve (AUC)) was 6190 h.ng/mL, the clearance moderate (CL = 3.1 L/h/kg) and the volume of
distribution was large (Vss = 4.2 L/kg). A rapid and sustained uptake of 12 in tumor tissue was observed,
associated with a slower decrease of compound 12 concentration from tumor tissue compared to the
plasma concentration. These results indicate a higher tumor to plasma exposure of 12 (with a tumor to
plasma AUC ratio equal to 8.1; Figure 6A).
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
Pharmacodynamic analysis in tumor tissue demonstrated a timeꢀdependent reduction of MET kinase
activity. Following administration of 12 at 20 mg/kg , phosphoꢀMET(Y1349) evaluation in tumor
extracts revealed that MET kinase inhibition was complete at 4 hours (96%), significant at 24 hours
(80%) and partial at 48 hours (61%) (Figure 6B). A direct relationship was observed between PD
modulation and tumor concentration of 12 rather than with plasma concentration in this murine model.
Significant PꢀMET inhibition at 24h was associated with compound concentration of about 300 ng/ml in
the tumor which represented an unbound drug exposure of about 2nM above the IC50 estimated in vitro
vs Hs 746T cell lines (1 nM). It is worth mentioning that high protein binding of 12 in plasma (99.67%)
did not prevent the molecule from distributing to the tumor tissue. This pharmacodynamic impact
translated into tumor regression, as previously described.20
ACS Paragon Plus Environment
12

Page 13 of 22
Journal of Medicinal Chemistry
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
Figure 6: Pharmacokinetic/Pharmacodynamic (PK/PD) study of compound 12 in mice bearing xenograft tumor of the METꢀ
amplified Hs 746T human gastric tumor cell line. Mice bearing established subcutaneous tumors derived from Hs 746T
gastric cancer cell line were administered a single 20 mg/kg dose of 12 IV in the tail vein. The pharmacokinetic parameters
of 12 were assessed by analysis of the drug concentrations in plasma and tumor tissue (A). The pharmacodynamic effect on
MET kinase was assessed by analysis of phosphoꢀMET(Y1349) levels in tumor extracts using Meso Scale Discovery (MSD)
immunoassay (B).
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
The PK parameters of 12 were further characterized at lower intravenous doses in mice, rats and dogs
(Table 3). In all 3 species evaluated the clearance was moderate. The plasma terminal elimination halfꢀ
life (t1/2) was moderate (rats and dogs) to long (mice). The volume of distribution at steady state was
moderate in dogs and large in rodents. The potential large tissue distribution in rodents was further
confirmed by performing a tissue distribution study in rats, with an infusion of 25 mg/kg [14C]ꢀ12.
Radioactivity was widely distributed throughout the whole body with maximum radioactivity
concentrations reached in most tissues at the end of the infusion (i.e., 1 hour) or 2 hours after the start of
infusion. Taken together, these results indicate a large distribution into the tumor tissue.
Table 3. Main PK parameters of 12 given IV in preꢀclinical models
Parametera
Mouse
Rat
Dog
Dose, mg/kg (IV)
1
2.8
54
3.2
4.8
5
1.5
36
2.9
3.6
3
0.5
20
1.3
3.8
CL (L/h/kg)b
CL as % of blood flow
Vss (L/kg)
t1/2 (h)
a Mean PK parameters in 3 female (mice and dogs) or male (rats) animals
b Plasma CL for animal species (blood /plasma ratio = 0.86)
ACS Paragon Plus Environment
13

Journal of Medicinal Chemistry
Page 14 of 22
Preliminary PK analysis from the Phase I clinical trial31 indicated that compound 12 blood exposure
(AUC) increased proportionally from 50 to 740 mg/m². After the first administration, the
concentrations decreased over time with a terminal halfꢀlife of 15.9 hours. Compound 12 exhibited a
medium blood clearance (mean 33.9 L/h corresponding to approximately 0.5 L/h/kg, coefficient of
variation (CV) 38%), representing approximately 37% of the hepatic blood flow (87 L/h) and a large
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
volume of distribution at steady state (mean 505 L corresponding to approximately 7.2 L/kg, CV 99%).
Taken together, the disposition of 12 was assessed in a range of animal species, and the distribution and
elimination processes documented in animals are consistent with the disposition observed in humans.
Thus, the animal pharmacological models tested were likely to be predictive for efficacy of 12 in
patients.
In conclusion, the chemical optimization of a focusedꢀscreen hit, supported by structural analysis, led to
the identification of compound 12, a very potent and selective MET kinase inhibitor with favorable
druggability properties for intravenous administration, and sustained target engagement in a METꢀ
amplified tumor model. Overall, the compound profile being compatible with a weekly schedule of
administration, further development with IV route was initiated, since some patients (e.g. METꢀ
amplified esophagogastric adenocarcinoma) could prefer an IV infusion to an oral tablet. It is also
possible that a less frequent dosing (weekly) versus a repeated format (daily) would trigger less acquired
resistance to such treatment. A favorable tolerance profile was observed in Phase I patients, with a dose
escalation performed up to the MAD of 740 mg/m2 (MTD 570 mg/m2). Early evidence of antitumor
activity triggered the evaluation of compound 12 in a Phase II clinical trial in METꢀamplified NSCLC
patients (NCT02435121).
ASSOCIATED CONTENT
Supporting Information
3D views presented here were generated using BIOVIA Discovery Studio 4.5/2016 software and
Cambridge Crystallographic Data Centre Mercury CFC 3.7. The atomic coordinates and structure
ACS Paragon Plus Environment
14

Page 15 of 22
Journal of Medicinal Chemistry
factors have been deposited in the Protein Data Bank, www.pdb.org; PDB ID codes: 5HLW (cpd 14),
5HNI (cpd 3), 5HO6 (SAR125844ꢀY1230H), 5HOA (SAR125844ꢀL1195V) and 5HOR (SAR125844ꢀ
M1250T).
1
2
3
4
5
6
7
8
9
AUTHOR INFORMATION
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
Corresponding Author
* Eꢀmail: laurent.schio@sanofi.com. Phone: +33(0)1 58 93 88 14.
Present Address
13 Quai Jules Guesde, 94403 VitryꢀsurꢀSeine Cedex, France
Notes
The authors declare no competing financial interest.
ACKNOWLEDGMENTS
This article is dedicated to the memory of our colleagues JeanꢀYves Crenne µ and Dominique Damour†.
The authors would like to thank Anne Dagallier‡ and Hélène de Foucauld‡ for modeling studies and
Jacques Dumas µ and Francis Duffieux µ for proofreading.
ABBREVIATIONS USED
HGF, Hepatocyte Growth Factor; NSCLC, NonꢀSmall Cell Lung Cancer; CRC, Colorectal Cancer;
PK/PD, Pharmacokinetic/Pharmacodynamic; DFG, AspꢀPheꢀGly; ITC, Isothermal Titration
Calorimetry; HPLC, High Performance Liquid Chromatography; MW, Molecular Weight; AUC, Area
Under the Curve; MSD, Meso Scale Discovery; CV, coefficient of variation
REFERENCES:
(1) Trusolino, L.; Bertotti A.; Comoglio PM. MET signaling: principles and functions in development,
organ regeneration and cancer. Nat. Rev. Mol. Cell Biol. 2010, 11, 834ꢀ848.
ACS Paragon Plus Environment
15

Journal of Medicinal Chemistry
Page 16 of 22
(2) Guerardi E.; Birchmeier W.; Birchmeier C.; Vande Woude G. Targeting MET in cancer: rationale
and progress. Nature Reviews Cancer 2012, 12, 89ꢀ103.
1
2
3
4
5
6
7
8
(3) Bladt, F.; Riethmacher, D.; Isenmann, S.; Aguzzi, A.; Birchmeier, C. Essential role for the cꢀmet
receptor in the migration of myogenic precursor cells into the limb bud. Nature 1995, 376, 768ꢀ771.
(4) Huh, C.G.; Factor, V.M.; Sánchez, A.; Uchida, K.; Conner, E.A.; Thorgeirsson, S.S. Hepatocyte
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
growth factor/cꢀmet signaling pathway is required for efficient liver regeneration and repair. Proc. Natl.
Acad. Sci. U S A 2004, 101(13), 4477ꢀ4482.
(5) Borowiak, M.; Garratt, A.N.; Wüstefeld, T.; Strehle, M.; Trautwein, C.; Birchmeier, C. Met provides
essential signals for liver regeneration. Proc. Natl. Acad. Sci. U S A 2004, 101(29), 10608ꢀ10613.
(6) Schmidt, L.; Duh, FM.; Chen, F.; Kishida, T.; Glenn, G.; Choyke, P.; Scherer, SW.; Zhuang, Z.;
Lubensky, I.; Dean, M.; Allikmets, R.; Chidambaram, A.; Bergerheim, UR.; Feltis, JT.; Casadevall, C.;
Zamarron, A.; Bernues, M.; Richard, S.; Lips, CJ.; Walther, MM.; Tsui, LC.; Geil, L.; Orcutt, ML.;
Stackhouse, T.; Lipan, J.; Slife, L.; Brauch, H.; Decker, J.; Niehans, G.; Hughson, MD.; Moch, H.;
Storkel, S.; Lerman, MI.; Linehan, WM.; Zbar, B. Germline and somatic mutations in the tyrosine
kinase domain of the MET protoꢀoncogene in papillary renal carcinomas. Nature Genetics 1997; 16(1),
68ꢀ73.
(7) KongꢀBeltran, M.; Seshagiri, S.; Zha, J.; Zhu, W.; Bhawe, K.; Mendoza, N.; Holcomb, T.; Pujara,
K.; Stinson, J.; Fu, L.; Severin, C.; Rangell, L.; Schwall, R.; Amler, L.; Wickramasinghe, D.; Yauch, R.
Somatic mutations lead to an oncogenic deletion of Met in lung cancer. Cancer Res. 2006, 66(1), 283ꢀ
289.
(8) Capuzzo, F.; Marchetti, A.; Skokan, M.; Rossi, E.; Gajapathy, S.; Felicioni, L.; Del Grammastro,
M.; Sciarrotta, MG.; Buttitta, F.; Incarbone, M.; Toschi, L.; Finocchiaro, G.; Destro, A.; Terracciano,
L.; Roncalli, M.; Alloisio, M.; Santoro, A.; VarellaꢀGarcia, M. Increased MET gene copy number
negatively affects survival of surgically resected nonꢀsmall cell lung cancer patients. J. Clin. Oncol.
2009, 27(10), 1667ꢀ1674.
ACS Paragon Plus Environment
16

Page 17 of 22
Journal of Medicinal Chemistry
(9) Engelman, J.A.; Zejnullahu, K.; Mitsudomi, T.; Song, Y.; Hyland, C.; Park, J.O. ; Lindeman, N. ;
Gale, CM.; Zhao, X. ; Christensen, J. ; Kosaka, T. ; Holmes, AJ.; Rogers, AM.; Cappuzzo, F. ; Mok,
T. ; Lee, C. ; Johnson, BE.; Cantley, LC. ; Jänne, PA. MET amplification leads to gefitinib resistance in
lung cancer by activating ERBB3 signaling. Science 2007, 316, 1039ꢀ1043.
1
2
3
4
5
6
7
8
9
(10) Bean, J.; Brennan, C.; Shih, J.Y.; Riely, G.; Viale, A.; Wang, L.; Chitale, D.; Motoi, N.; Szoke, J.;
Broderick, S.; Balak, M.; Chang, WC.; Yu, CJ.; Gazdar, A.; Pass, H.; Rusch, V.; Gerald, W.; Huang,
SF.; Yang, PC.; Miller, V.; Ladanyi, M.; Yang, CH.; Pao, W. MET amplification occurs with or without
T790M mutations in EGFR mutant lung tumors with acquired resistance to gefitinib or erlotinib. Proc.
Natl. Acad. Sci. U S A 2007, 104, 20932ꢀ20937.
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
(11) Sequist, L.V.; Waltman, B.A.; DiasꢀSantagata, D.; Digumarthy, S.; Turke, A.B.; Fidias, P.;
Bergethon, K.; Shaw, AT.; Gettinger, S.; Cosper, AK.; Akhavanfard, S.; Heist, RS.; Temel, J.;
Christensen, JG.; Wain, JC.; Lynch, TJ.; Vernovsky, K.; Mark, EJ.; Lanuti, M.; Iafrate, AJ.; Minoꢀ
Kenudson, M.; Engelman, JA. Genotypic and histological evolution of lung cancers acquiring resistance
to EGFR inhibitors. Sci. Transl. Med. 2011, 3, 75ra26.
(12) Bardelli, A.; Corso, S.; Bertotti, A.; Hobor, S.; Valtorta, E.; Siravegna, G.; SartoreꢀBianchi, A.;
Scala, E.; Cassingena, A.; Zecchin, D.; Apicella, M.; Migliardi, G.; Galimi, F.; Lauricella, C.; Zanon,
C.; Perera, T.; Veronese, S.; Corti, G.; Amatu, A.; Gambacorta, M.; Diaz, LA;. Sausen, M.; Velculescu,
VE.; Comoglio, P.; Trusolino, L.; Di Nicolantonio, F.; Giordano, S. ; Siena, S. Amplification of the
MET receptor drives resistance to antiꢀEGFR therapies in colorectal cancer. Cancer Discovery 2013, 3,
658ꢀ673.
(13) Straussman, R.; Morikawa, T.; Shee, K.; BarzilyꢀRokni, M.; Qian, Z.R.; Du, J.; Davis, A.;
Mongare, M.M.; Gould, J.; Frederick, D.T.; Cooper, Z.A.; Chapman, P.B.; Solit, D.B.; Ribas, A.; Lo,
R.S.; Flaherty, K.T.; Ogino, S.; Wargo, J.A.; Golub, T.R. Tumour microꢀenvironment elicits innate
resistance to RAF inhibitors through HGF secretion. Nature 2012, 487, 500ꢀ504.
(14) Liu, X.; Wang, Q.; Yang, G.; Marando, C.; Koblish HK.; Hall, LM.; Fridman, JS.; Behshad, E.;
Wynn, R.; Li, Y.; Boer, J.; Diamond, S.; He, C.; Xu, M.; Zhuo, J.; Yao, W.; Newton, RC.; Scherle, PA.
ACS Paragon Plus Environment
17

Journal of Medicinal Chemistry
Page 18 of 22
A novel kinase inhibitor, INCB28060, blocks cꢀMETꢀdependent signaling, neoplastic activities, and
crossꢀtalk with EGFR and HERꢀ3. Clin. Cancer Res. 2011, 17, 7127ꢀ7138.
1
2
3
4
5
6
7
8
(15) Bladt, F.; Faden, B.; FrieseꢀHamim, M.; Knuehl, C.; Wilm, C.; Fittschen, C.; Gradler, U.; Meyring,
M.; Dorsch, D.; Jaehrling, F.; Pehl, U.; Stieber, F.; Schadt, O.; and Blaukat, A. EMD 1214063 and
EMD 1204831 constitute a new class of potent and highly selective cꢀMet inhibitors. Clin. Cancer Res.
2013, 19, 2941ꢀ2951.
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
(16)Jia, H.; Dai, G.; Weng, J.; Zhang, Z.; Wang, Q.; Zhou, F.; Jiao, L.; Cui, Y.; Ren, Y.; Fan, S.; Zhou,
J.; Qing, W.; Gu, Y.; Wang, J.; Sai, Y.; Su, W. Discovery of (S)ꢀ1ꢀ(1ꢀ(imidazo[1,2ꢀ a ]pyridinꢀ6ꢀ
yl)ethyl)ꢀ6ꢀ(1ꢀmethylꢀ1 H ꢀpyrazolꢀ4ꢀyl)ꢀ1 H ꢀ[1,2,3]triazolo[4,5ꢀ b ]pyrazine (volitinib) as a highly
potent and selective mesenchymalꢀepithelial transition factor (cꢀMet) inhibitor in clinical development
for treatment of cancer. J. Med. Chem. 2014, 57, 7577ꢀ7589.
(17) Hugues, PE.; Yang, Y.; Rex, K.; Zhang, Y.; KaplanꢀLefko, PJ.; Caenepeel, S.; Moriguchi, J.;
Broome, M.; Choquette, D.; Radinsky, R.; Kendall, R.; Coxon, A.; Dussault, I. AMG 337, a novel,
potent and selective MET kinase inhibitor, has robust growth inhibitory activity in METꢀdependent
cancer models. Cancer Res. 2014, 74(19 Suppl): Abstract 728.
(18) Okamoto, W.; Okamoto, I.; Arao, T.; Kuwata, K.; Yamaguchi, H.; Sakai, K.; Yanagihara, K.;
Nishio, K.; Nakagawa, K. Antitumor action of the MET tyrosine kinase inhibitor Crizotinib (PFꢀ
02341066) in gastric cancer positive for MET amplification. Mol. Cancer Ther. 2012, 11, 1557ꢀ1564.
(19) Schio, L.; Nemecek, C.; Ugolini, T.; Wentzler, S.; Grapinet, S.; Khider, J.; Albert, E.; Dischamps,
N.; Gay, F.; Sonnefraud, V.; Bacqué, E.; Kenigsberg, M.; Goulaouic, H.; Dagallier, A.; Vallée, F.;
Bonche, F.; Lengauer, C. SAR125844: a potent and selective ATPꢀcompetitive inhibitor of MET kinase.
Cancer Res. 2012, 72(8 Suppl 1): Abstract 2911.
(20) Egile, C.; Kenigsberg, M.; Delaisi, C.; Begassat, F.; DoꢀVale, V.; Mestadier, J.; Bonche, F.;
Benard, T.; Nicolas, J.P.; Valence, S.; Lefranc, C.; Francesconi, E.; Castell, C.; Lefebvre, A.M.;
Nemecek, C.; Calvet, L.; Goulaouic, H. The selective intravenous inhibitor of the MET tyrosine kinase
SAR125844 inhibits tumor growth in METꢀamplified cancer. Mol. Cancer Ther. 2015, 14, 384-394.
ACS Paragon Plus Environment
18

Page 19 of 22
Journal of Medicinal Chemistry
(21) Zhao, Z.; Wu, H.; Wang, L.; Liu, Y.; Knapp, S.; Liu, Q.; Gray N. S. Exploration of type II binding
mode: a privileged approach for kinase inhibitor focused drug discovery. ACS Chem. Biol. 2014, 9,
1230ꢀ1241.
1
2
3
4
5
6
7
8
9
(22) a).Boezio, A.A.; Copeland, K.W.; Rex, K.; Albrecht, B.K.; Bauer, D.; Bellon, S.F.; Boezio, C.;
Broome, M.A.; Choquette, D.; Coxon, A.; Dussault, I.; Hirai, S.; Lewis, R.; Lin, MꢀH.J.; Lohman, J.;
Liu, J.; Peterson, E.A., Potashman, M.; Shimanovich, R.; Teffera, Y.; Whittington, D.A.; Vaida, K.R.;
Harmange, JꢀC. Discovery of (R)‑6ꢀ(1ꢀ(8ꢀfluoroꢀ6ꢀ(1ꢀmethyl‑1H‑pyrazolꢀ4ꢀyl)ꢀ[1,2,4]triazolo[4,3‑
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
a]pyridinꢀ3ꢀyl)ethyl)ꢀ3ꢀ(2ꢀethoxyethoxy)ꢀ1,6ꢀnaphthyridinꢀ5(6H)‑one (AMG 337), a potent and
selective inhibitor of MET with high unbound target coverage and robust in vivo antitumor activity. J.
Med.Chem. 2016, 59(6), 2328ꢀ2342. b). Lolkema, M.P.; Bohets, H.H.; Arkenau, H.T.; Lampo, A.;
Barale, E.; de Jonge, M.J.; van Doorn, L.; Hellemans, P.; de Bono, J.S.; Eskens, F.A. The cꢀMet
tyrosine kinase inhibitor JNJꢀ38877605 causes renal toxicity through speciesꢀspecific insoluble
metabolite formation. Clin. Cancer Res. 2015, 21(10), 2297ꢀ304.c). Buchanan, S.G.; Hendle, J.; Lee,
P.S.; Smith, C.R.; Bounaud, P.Y.; Jessen, K.A.; Tang, C.M.; Huser, N.H.; Felce, J.D.; Froning, K.J.;
Peterman, M.C.; Aubol, B.E.; Gessert, S.F.; Sauder, J.M.; Schwinn, K.D.; Russell, M.; Rooney, I.A.;
Adams, J.; Leon, B.C.; Do, T.H.; Blaney, J.M.; Sprengeler, P.A.; Thompson, D.A.; Smyth, L.; Pelletier,
L.A.; Atwell, S.; Holme, K.; Wasserman, S.R.; Emtage, S.; Burley, S.K.; Reich, S.H. SGX523 is an
exquisitely selective, ATPꢀcompetitive inhibitor of the MET receptor tyrosine kinase with antitumor
activity in vivo. Mol. Cancer Ther. 2009, 8(12), 3181ꢀ90.
(23) Kawasaki, Y.; Freire E. Finding a better path to drug selectivity. Drug Discovery Today, 2011, 16,
985-990.
(24) Bissantz, C.; Kuhn, B.; Stahl, M. A medicinal chemist’s guide to molecular interactions. J. Med.
Chem. 2010, 53, 5061-5084.
ACS Paragon Plus Environment
19

Journal of Medicinal Chemistry
Page 20 of 22
(25) Kornev, A.P; Nina M; Haste, N.M.; Taylor, S.S.; Eyck, L.F.T. Surface comparison of active and
1
2
3
4
5
inactive protein kinases identifies a conserved activation mechanism. Proc. Natl. Acad. Sci., 2006, 107,
17783-17788.
6
7
8
9
(26) Novel 6ꢀtriazolopyridazinesulfanyl benzothiazole and benzimidazole derivatives, method for
production thereof and application as medicaments and pharmaceutical compositions and novel use as
MET inhibitors (Applicant: SanofiꢀAventis Fr.). Eva Albert, Eric Bacque, Conception Nemecek,
Antonio Ugolini, Sylvie Wentzler. WO2009056692 A2; May 7, 2009.
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
(27) Leeson, P.D.; Young, R.J. Molecular property design: does everyone get it? ACS Med. Chem. Lett.
2015, 6, 722-725.
(28) Veber, D. F.; Johnson, S. R.; Cheng, H.ꢀY.; Smith, B. R.; Ward, K. W.; Kopple, K.D. Molecular
properties that influence the oral bioavailability of drug candidates. J. Med.Chem. 2002, 45, 2615ꢀ2623.
(29) Doak, B. C.; Zheng, J.; Dobritzsch, D.; Kihlberg, J. How beyond rule of 5 drugs and clinical
candidates bind to their targets. J. Med.Chem. 2016, 59(6), 2312ꢀ2327.
(30) a). Derivatives of 6ꢀ(6ꢀOꢀcycloalkyl or 6ꢀNHꢀcycloalkylꢀtriazolopyridazineꢀsulfanyl)
benzothiazoles and benzimidazoles, preparation thereof, use thereof as drugs, and use thereof as MET
inhibitors. (Applicant: SanofiꢀAventis Fr.); Eric Bacque, Conception Nemecek, Antonio Ugolini, Sylvie
Wentzler; WO2010089506 A1; Aug 12, 2010. b). Derivatives of 6ꢀ(6ꢀOꢀsubstitutedꢀtriazolopyridazineꢀ
sulfanyl) benzothiazoles and benzimidazoles, preparation thereof, use thereof as drugs, and use thereof
as MET inhibitors (Applicant: SanofiꢀAventis Fr.); Conception Nemecek, Antonio Ugolini, Sylvie
Wentzler; WO2010089507 A1; Aug 12, 2010. c). Derivatives of 6ꢀ(6ꢀNHꢀsubstitutedꢀ
triazolopyridazineꢀsulfanyl) benzothiazoles and benzimidazoles, preparation thereof, use thereof as
drugs, and use thereof as MET inhibitors (Applicant: SanofiꢀAventis Fr.); Eric Bacque, Conception
Nemecek, Antonio Ugolini, Sylvie Wentzler; WO2010089508 A1; Aug 12, 2010.
(31) Angevin, E.; Spitaleri, G.; Hollebecque, A.; De Pas, T.; Soria, J.C.; Harnois, M.; Mazuir, F. ;
Assadourian, S.; De Marinis, F. A firstꢀinꢀhuman (FIH) phase I study of SAR125844, a novel selective
ACS Paragon Plus Environment
20

Page 21 of 22
Journal of Medicinal Chemistry
MET kinase inhibitor, in patients (pts) with advanced solid tumors: Dose escalation results. J. Clin.
Oncol. 2014, 32(5 Suppl): Abstract 2506.
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
ACS Paragon Plus Environment
21

Journal of Medicinal Chemistry
Page 22 of 22
1
2
Graphic for Table of Contents Only:
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
ACS Paragon Plus Environment
22