Design and synthesis of novel selective anaplastic lymphoma kinase inhibitors

Article abstract of DOI:10.1016/j.bmcl.2015.11.049

Anaplastic lymphoma kinase (ALK) is a receptor tyrosine kinase belonging to the insulin receptor superfamily. Expression of ALK in normal human tissues is only found in a subset of neural cells, however it is involved in the genesis of several cancers through genetic aberrations involving translocation of the kinase domain with multiple fusion partners (e.g., NPM-ALK in anaplastic large cell lymphoma ALCL or EML4-ALK in non-small cell lung cancer) or activating mutations in the full-length receptor resulting in ligand-independent constitutive activation (e.g., neuroblastoma). Here we are reporting the discovery of novel and selective anaplastic lymphoma kinase inhibitors from specific modifications of the 2,4-diaminopyridine core present in TAE684 and LDK378. Synthesis, structure activity relationships (SAR), absorption, distribution, metabolism, and excretion (ADME) profile, and in vivo efficacy in a mouse xenograft model of anaplastic large cell lymphoma are described.

Full text of DOI:10.1016/j.bmcl.2015.11.049

Accepted Manuscript
Design and Synthesis of Novel Selective Anaplastic Lymphoma Kinase Inhib-
itors
Pierre-Yves Michellys, Bei Chen, Tao Jiang, Yunho Jin, Wenshuo Lu, Thomas
H. Marsilje, Wei Pei, Tetsuo Uno, Xuefeng Zhu, Baogen Wu, Truc Ngoc
Nguyen, Badry Bursulaya, Christian Lee, Nanxin Li, Sungjoon Kim, Tove
Tuntland, Bo Liu, Frank Sun, Auzon Steffy, Tami Hood
PII:
S0960-894X(15)30257-2
DOI:
http://dx.doi.org/10.1016/j.bmcl.2015.11.049
BMCL 23305
Reference:
Bioorganic & Medicinal Chemistry Letters
To appear in:
Received Date:
Revised Date:
Accepted Date:
3 August 2015
12 November 2015
16 November 2015
Please cite this article as: Michellys, P-Y., Chen, B., Jiang, T., Jin, Y., Lu, W., Marsilje, T.H., Pei, W., Uno, T.,
Zhu, X., Wu, B., Nguyen, T.N., Bursulaya, B., Lee, C., Li, N., Kim, S., Tuntland, T., Liu, B., Sun, F., Steffy, A.,
Hood, T., Design and Synthesis of Novel Selective Anaplastic Lymphoma Kinase Inhibitors, Bioorganic &
Medicinal Chemistry Letters (2015), doi: http://dx.doi.org/10.1016/j.bmcl.2015.11.049
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.

Graphical Abstract
To create your abstract, type over the instructions in the template box below.
Fonts or abstract dimensions should not be changed or altered.
Design and Synthesis of Novel Selective Anaplastic Lymphoma Kinase
Inhibitors
Leave this area blank for abstract info.
Pierre-Yves Michellys, Bei Chen Tao Jiang, Yunho Jin, Wenshuo Lu, Thomas H. Marsilje, Wei Pei, Tetsuo Uno, Xuefeng Zhu, Baogen Wu, Truc Ngoc
Nguyen, Badry Bursulaya, Christian Lee, Nanxin L, Sungjoon Kim, Tove Tuntland, Bo Liu, Frank Sun, Auzon Steffy and Tami Hood

Bioorganic & Medicinal Chemistry Letters
journal homepage: www.els evier.com
Design and Synthesis of Novel Selective Anaplastic Lymphoma Kinase Inhibitors
Pierre-Yves Michellysa,*, Bei Chena Tao Jianga, Yunho Jina, Wenshuo Lua, Thomas H. Marsiljea, Wei Peia,
Tetsuo Unoa, Xuefeng Zhua, Baogen Wua, Truc Ngoc Nguyena, Badry Bursulayaa, Christian Leea, Nanxin
Lia, Sungjoon Kima, Tove Tuntlanda, Bo Liua, Frank Sunb, Auzon Steffya and Tami Hoodc
a Genomics Institute of the Novartis Research Foundation, 10675 John Jay Hopkins Drive, San Diego, CA 920121
* To whom correspondence should be addressed
b Sanofi Oncology, 640 Memorial Drive, Cambridge, MA 02139
c Novartis Institutes for BioMedical Research, Inc., 250 Massachusetts Avenue, Cambridge, MA 02139
ARTICLE INFO
ABSTRACT
Article history:
Received
Anaplastic lymphoma kinase (ALK) is a receptor tyrosine kinase belonging to the insulin
receptor superfamily. Expression of ALK in normal human tissues is only found in a subset of
neural cells, however it is involved in the genesis of several cancers through genetic aberrations
involving translocation of the kinase domain with multiple fusion partners (e.g. NPM-ALK in
anaplastic large cell lymphoma ALCL or EML4-ALK in non-small cell lung cancer ) or
activating mutations in the full-length receptor resulting in ligand-independent constitutive
activation (e.g neuroblastoma). Here we are reporting the discovery of novel and selective
anaplastic lymphoma kinase inhibitors from specific modifications of the 2,4-diaminopyridine
core present in TAE684 and LDK378. Synthesis, structure activity relationships (SAR),
absorption, distribution, metabolism, and excretion (ADME) profile, and in vivo efficacy in a
mouse xenograft model of anaplastic large cell lymphoma are described.
Revised
Accepted
Available online
Keywords:
ALK
Scaffold morphing
Hinge
Karpas299
2009 Elsevier Ltd. All rights reserved
.

Anaplastic lymphoma kinase (ALK) is a receptor tyrosine
kinase of the insulin receptor superfamily and expression of
ALK in normal human tissues is only found in a subset of
neural cells.1 It is involved however in the genesis of several
cancers through genetic aberrations involving translocation of
the kinase domain with multiple fusion partners or activating
mutations that result in ligand-independent constitutive
activation.2-4 To date, no essential role has been found for
ALK in mammals. Mice deficient in ALK have normal
development and display an anti-depressive profile with
enhanced performance in hippocampus-dependent tasks
potentially due to increased hippocampal progenitor cells.5
within reach of the carbonyl from the Glu1197 towards the
gatekeeper region (Figure 2). This conformation was
confirmed later on when a co-crystal was obtained.26 We
postulated it was likely feasible to introduce an extra
interaction with Glu1197 by a simple addition of a hydrogen
donor in this region. This would result into a hinge binding
moieties containing three donor-acceptor interactions instead
of two in the pyrimidine case.
Deregulation of ALK was first identified in anaplastic large
cell lymphoma (ALCL) where the tyrosine kinase domain is
fused to nucleophosmin (NPM), a product of recurrent
t(2;5)(p23;q35) chromosome translocation.6 Subsequently,
chromosome rearrangements resulting in ALK fused to
various partner genes have been found in nearly 70% of
ALCL, 40-60% of inflammatory myofibroblastic tumors
(IMT),7 a few dozen cases of diffuse large B-cell lymphoma
(DLBCL), and most recently in 2-7% of non-small cell lung
cancer (NSCLC).8-10 Among fusion partner genes identified to
date, NPM is the most common partner in ALCL and
echinoderm microtubule-associated protein-like-4 (EML4) is
the main partner in NSCLC. In addition to the chromosome
rearrangements that result in ALK fusion genes, amplification
of ALK gene and activating point mutations in the full length
ALK gene have recently been reported in neuroblastoma,11-13
inflammatory breast cancer,14 and ovarian cancer.15
Figure 2. Co-crystal structure of 2 complexed with the
ALK kinase domain depicting the proximity of Glu1197 to the
pyrimidine ring forming interactions with Met1199 at the hinge
moiety.27
In the aim to conduct our studies (and further test our
hypothesis), we decided to include some ring systems that lack
the possibility to involve a third interaction at the hinge like a
quinazoline (F) and a pyrimidopyrimidine (G) in addition to
the 5,6-fused ring systems like a pyrrolopyrimidine (A), a
purine (B), or a pyrrazolopyrimidine (D). The ring systems
and the anilines I1-I3 we used to make the final molecules are
depicted in Figure 3.
To date, 3 (crizotinib, Xalkori®)16, 17 has been approved for
the treatment ALK-positive NSCLC and 2 (LDK378, ceritinib,
Zykadia®)18 was approved for the treatment of crizotinib-
resistant NSCLC patients. Both 4 (alectinib) and 5 (AP26113)
have obtained the breakthrough therapy designation by the
FDA for their activity in crizotinib-resistant NSCLC patients.
Figure 1 represents a selected subset of ALKi currently FDA
approved or in clinical trials. 18-25
Figure 3. Rings systems (A-H) and anilines (I1-I3) used to
make the final molecules.
The syntheses of the compounds described in this
communication (11a-e, 12a-e, 16a,b, 20a,b, 23, 26, 27, 30a,b,
31 and 32) mirror the synthetic route we described for the
synthesis of 218 and are depicted in schemes 1-4. For the sake
of clarity, we have voluntarily restricted the analogues shown
in this communication as ones bearing an unsubstituted
piperidine (compounds 11a-e, 16a, 20a, 31 and 33) or N-Me-
substituted piperidine ring (compounds 12a-e, 16b, 20b, 23,
26, 27 and 30a,b). Typically, we introduced the 2-(iso-
propylsulfonyl)amino moiety first using simple amination
conditions (usually, IPA, reflux) by condensation of 9a-e, 14,
17, 21 and 28a-c with 2-(iso-propylsulfonyl)aniline to afford
10a-e, 15, 18, 22 and 29a-c in moderate to excellent yields. In
the case of compounds 11a-e, 30a,b, 29a-c and 31 the
synthesis was completed by a second amination reaction using
selected proprietary aniline derivatives (I1-I3, Figure 3) in
moderate to good yields.18,28 In the case of compounds 16a,b,
and 23, an additional synthetic step was introduced on 18 and
22 in order to form the pyrrazolopyrimidine ring
(NH2NH2.2HCl, NaOAc, EtOH, 80oC). For compounds 16a,
26 and 27, an additional deprotection step of the tosylate and
Figure 1. Selected examples of ALKi.
In
a
previous communication, we presented the
modifications we made around TAE684 (1) that led to the
discovery of LDK378 (2).18 We present additional medicinal
chemistry efforts performed on this scaffold; focusing in this
communication on the replacement of the pyrimidine ring
present in compound 2 in the aim of optimizing hinge
interactions.
2 binds to the hinge of the ALK kinase via interactions of
the N1 of the pyrimidine ring and N2 from the aniline bearing
the solubilizing moiety.26 As we described previously, docking
of 2 in the ALK kinase domain revealed the aminopyrimidine
making contact at the hinge with the Met1199 residue and was

carbamate groups was necessary to complete the synthesis.
Further alkylation (typically, CH3-I, Et3N, DMF, MW, 100oC,
10 minutes) of compounds 11a-e afforded the derivatives 12a-
e in good yields (>70%). Reductive amination (HCHO,
MeOH/THF, NaBH3CN) of 16a yielded compound 16b.
Compounds 26 and 27 required an inverted sequence of
reactions in order to be synthesized (amination using 2-(iso-
propylsulfonyl)aniline failed to produce the desired derivative,
likely due to the lower reactivity of the aniline). Their
synthesis was achieved by two sequential Buchwald couplings
involving at first the aniline I3 and at second the 2-(iso-
propylsulfonyl)amino moiety. Finally, reduction of the
pyridine ring (PtO2, AcOH, TFA) of 31 yielded 32 in good
yield.
Scheme 3. Synthesis of derivatives 26 and 27. Reagents
and conditions: a. Pd (OAc)2, Xantphos, C2CO3, THF, MW,
150oC, 40 min. b. I3, Pd2(dba)3, X-Phos, NaotBu, THF, MW,
150oC, 40 min.
Scheme 4. Synthesis of derivatives 30a,b, 31 and 32.
Reagents and conditions: a. 2-(iso-propylsulfonyl)aniline,
conc. HCl, dioxane, 150oC MW.
b. I3, conc. HCl,
CH3CH2CH2OH, 160oC, MW, 20 min. d. H2, PtO2, AcOH,
TFA.
As described in our previous communication,18 ALK
inhibition was directly measured in a cellular context by
measuring the proliferation of Ba/F3 cells expressing NPM-
ALK as a guide for our SAR. Ba/F3 cells expressing the Tel-
InsR fusion protein as well as wild-type (WT) Ba/F3 cells
were used as counter screens. From our studies, several
compounds 11a, 11d, 12a-e and 20a,b display IC50s below 50
nM (47, 33, 6, 15, 8, 13, 47, 38 and 20 nM respectively) while
eight compounds (16a,b, 26, 27, 30a,b, 31 and 32) suffer a
strong loss in potency (684, 1298, 1427, 1119, 1082, 1286,
3172 and 1326 nM respectively). The other compounds (11b,
11c and 23) were moderately potent against ALK (114, 66 and
150 nM respectively). We were pleased to see that 12a-d and
20b were potent ALK inhibitors with IC50s equal or below 20
nM (6, 15, 8, 13 and 20 nM respectively). From this study, it
appeared quickly that most 6,5-fused ring systems tried were
tolerated except when a substitution was introduced at the 2-
position of the pyrrolopyrimidine core (compounds 16a,b) or
when one of the nitrogen from the bicyclic system was
removed (compound 26 and 27).30 Typically, 6,6-fused ring
system were not well tolerated (30a,b, 31 and 32).30 The result
obtained with compound 31 was predictable due to the fact 31
contains an H-bond acceptor instead of an H-bond donor
Scheme 1. Synthesis of derivatives 11a-e, 12a-e, and
16a,b.
Reagents
and
conditions:
a.
2-(iso-
propylsulfonyl)aniline, conc. HCl, IPA, 150oC MW. b. NaH,
DMF/DMSO (10:1 v/v), 0oC to RT. c. I2, conc. HCl, IPA,
160oC, MW, 30 min. d. CH3-I, Et3N, DMF, MW, 100oC 10
min.
e.
Chloroacetone
(1.2 eq.), NaOAc (2.1 eq.), H2O, reflux, 3 hr. f. POCl3,
DIEPA, toluene, 70 to 110oC, O/N. g. (1) NaH, TsCl, DMF,
0oC, 1 hr. (2) 10% NH4Cl. h. NaOMe/MeOH, 50oC, 2 hrs then
RT O/N. i. HCHO (37%/H2O), MeOH/THF (1/1 v/v), RT, 2
hrs followed by NaBH3CN, RT, 30 min.
N
NH
Cl
O
Cl
N
N
Cl
N
OHC
HN
a
N
OHC
Cl
N
O
HN
N
NH
N
b, c
O
d, e, f
O
N
Cl
S
O
O
HN
Cl
O
S
Cl
O
S
17
18
19
N
R
20a
R = CH3, 20b
R = H,
N
NH
Cl
N
H2
N
N
Cl
N
NC
HN
N
NC
Cl
a
O
HN
N
NH
N
O
g, h
O
Cl
S
O
O
Cl
S
21
22
23
needed to form a hydrogen bond with the carbonyl of Glu1197.
N
The low potency with compound 32 is likely due to a larger
flexibility of the tetrahydropyridine moiety inducing a sub-
optimal positioning of the NH bond towards Glu1197. All
compounds show a moderate to good selectivity (20-76 fold)
versus InsR (12a having the highest selectivity ratio of 76
fold) except for compounds 16a,b, 26, 27, 30a,b, 31 and 32
where the selectivity was mostly abrogated likely due to the
sharp drop in potency for ALK. Typically, no significant
viability impairment of BaF3 cells was observed (BaF3-WT
column, table 1).
Scheme 2. Synthesis of derivatives 20a,b and 23. Reagents
and conditions: a. 2-(iso-propylsulfonyl)aniline, conc. HCl,
IPA, 150oC MW. b. CH3MgBr, THF, -78oC to RT. c. PDC,
DCM, RT. d. I2 or I3 conc. HCl, IPA, 160oC, MW, 30 min. e.
NH2NH2.2HCl, NaOAc, EtOH, 80oC. f. TFA/DCM. g. TFA,
CH2Cl2, RT. h. CH3-I, Et3N, DMF, MW, 100oC, 10 min. g. I3
conc. HCl, IPA, 160oC, MW, 30 min. h. NH2NH2, IPA, reflux

BaF3-
BaF3
BaF3-WT
Comp
ound
27
30b
32
2867 862
1344 405
6851 2311
186 10
1930 320
212 13
NMP-ALK
Tel-InsR
2
1
151 18
1643 272
319 24
700 100
2700 200
938 38
3479 180
2477 448
2216 100
>10
26
47
1
11a
11b
11c
11d
12a
12b
12c
12d
12e
16a
16b
20a
20b
23
3
Table 2. Activity of selected derivatives against the
Karpas299 cancer cell line and ALK enzyme.
114 27
66
33 20
9
3521 450
6220 1230
3422 251
3180 1870
4549 680
5120 520
>10000
1913 519
455 37
6
1
15
8
4
477 60
1
408 39
13
1
703 207
5058 780
4390 410
3933 560
1149 200
429 41
47 14
684 69
1298 152
5411 630
5731 760
5540 600
2507 747
4503 900
4912 550
2599 532
3240 170
3940 110
7230 700
2033 342
38
20
4
7
150 13
3451 970
5682 700
1666 281
2232 291
2240 460
5925 720
2656 1215
26
1427 151
1119 150
1082 132
1286 91
3172 350
1326 80
27
Figure 4. Co-crystal structure of 12d complexed to the
ALK kinase domain.29 A clear H-bond is visible between
GLU1197 and the hydrogen from the pyrrazolopyrimidine ring.
30a
30b
31
The co-crystal structure of 12d with ALK was obtained and
shown in Figure 4. There is a clear hydrogen bond between
the NH of the pyrrazolopyrimidine ring and the carbonyl of
32
GLU1197
.
This experimental structure confirmed our
Table 1. Activity profile of compounds 11a-d, 12a-e,
16a,b, 20a,b, 23, 26, 27, 30a,b, 31 and 32. All data given in
nM and are an average of duplicate measurements.
hypothesis and also correlated well with the SAR observed
with the compounds described.
A selected set of these derivatives were tested for their
ADME profile (metabolic clearance, CYP3A4 inhibition, HT-
solubility at pH 6.8, PAMPA, Caco2 and hERG binding). The
results are summarized in Table 3. All compounds displayed a
low to moderate microsomal clearance. CYP inhibition was
typically moderate (around 5 µM for most derivatives) and
ranged from about 3 µM (12d) to >25 µM (20b). A wide
range of solubility profiles were also observed (from 8 to >175
µM) which did not correlate well with the basicity of the
amine present on the piperidine ring. For example, 12a and
12b have the same N-methyl piperidine moiety (identical
calculated pKas of 9.48) but have a drastic different solubility
profile (10.4 versus >175 µM). Most compounds had good
permeability (PAMPA) except 11a. 11a and 12b-c showed an
efflux potential in the caco2 assay. hERG binding IC50s were
also quite widespread, ranging from 1 µM (11a and 12d) to
>30 µM (20b). Overall, compound 20b appeared to have a
superior profile compared to the other compounds listed in
Table 3 and proved to be very selective for ALK. When tested
enzymatically against a panel of 28 kinases (Table 3), 20b
displayed IC50 below 1 µM for only the following 3 kinases:
IGF-1R, CDK2 and HER1 (0.47, 0.92 and 0.96 µM
respectively).
Since we did observe a poorer activity in Ba/F3 cells for
some of the derivatives described in Table 1, we ran a subset
of these compounds in the Karpas299 cell line (patient cell
line harboring the NMP-ALK fusion) as well as an enzymatic
ALK assay in order to confirm their poor cellular potency was
due to a lower affinity for ALK and not due to other
parameters such as cell permeability. The results of this
selected set of compounds tested are described in Table 2.
These two assays show a strong correlation with each other
and also with the BaF3 results (moderately to highly potent
derivatives have similar IC50s on all assays (11b, 11c, 12a,b
and 20a,b) while weak derivatives from our Ba/F3 SAR
driving assay show a much weaker activity in both Karpas299
and ALK enzyme assay (16a, 26, 27, 30b and 32).
Compound
2
Karpas299
64
ALK enzyme
6
7
2
1
11b
11c
118 13
57 18
0.3
20 0.3
12a
12b
16a
20a
20b
26
3
6
0.8
1
8
2
0.2
0.2
Kinase
IC50
0.03
0.01
9.4
Kinase
EphB4
FGFR4
FGFR3
IC50
2.9
328 30
0.2
3878 1110
NPM-ALK
EML4-ALK
BTK
7
5
4
1.0
0.1
>10
>10
26 4.4
7310 1180
>25000

ABL
1.4
HER1
HER2
IGF-1R
KDR
0.96
7.8
AKT
7.6
Aurora
CDK2
GSK3β
JAK1
>10
0.92
4.3
0.47
7.4
LCK
4.3
>10
>10
>10
>10
3.9
PDGFRα
PDK1
RET
9.7
JAK2
>10
3.6
JAK3
MAPK1
PLK1
TYK2
EphA4
SYK
>10
>10
8.1
Figure 5. 20b activity in SCID mice Karpas299
xenograft.27
cKit
In conclusion, we successfully designed and synthesized
novel potent ALKi based on structural information available
from the 2,4-diaminopyrimidine hinge-binding moiety present
in ceritinib. The design was directed towards the morphing of
the pyrimidine ring contained in 2 into various bicyclic
systems that possess an additional interaction at the hinge with
Glu1197 allowed us to reach our goal in identifying potent
derivatives such as 12a-d or 20b. Compound 20b possesses an
attractive profile (potent on target, high selectivity against
other kinases, good solubility, low hERG inhibition, low CYP
inhibition), has an acceptable pharmacokinetic profile in mice
and rats and induces tumor regression in a Karpas299
xenograft mouse model of ALCL at 50 mg/kg when dosed
once a day.
>10
>10
MET
Table 3. Enzymatic profile of compound 20b. All data
given in nM and are an average of duplicate measurements.
Table 5 shows the in vivo mouse and rat PK profile of
compounds 11a, 12a-c and 20b (11a and 12b were not tested
for their rat PK due to their poor PK properties in mice). All
compounds display a moderate clearance in mice (40-71
mL/min/kg), and rats (15-32 mL/min/kg), they tend to have a
high Vss (7-19 L/kg) except 12b which had a much lower Vss
in mice (2.4 L/Kg). All compounds had a relatively low Cmax
in mice (29-429 nM) and rats (151-291 nM) and a moderate
bioavailability in both species (35-58%) which typically was
higher in mice, except for 9a and 10b which show a very poor
bioavailability in mice.
Based on these data, we decided to evaluate 20b in a two
week Karpas299 SCID mice xenograft model harboring the
NMP-ALK fusion (Figure 5). When dosed once a day at doses
of 25 and 50 mg/kg, 20b demonstrated a dose-dependent anti-
tumor activity with significant tumor regression at 50 mg/kg.31
Tumor changes were measured as %T/C where changes in
tumor weight for each treated (T) and control (C) groups were
measured and a percent of the ratio was calculated. Tumor
shrinkage (37% T/C) was observed at 25 mg/kg and tumor
regression (-54% T/C) was observed at 50 mg/kg. All doses
were well tolerated with no body weight loss (data not shown).

compound
CLint_m, r, h1
CYP3A42
HT-sol. 3
Log PAMPA4
Caco2
hERG5
(A-B/B-A)
0.04/1.3*
1.3/1.1
11a
12a
12b
12c
12d
20b
15, 14, 22
62, 34, 25
52, 28, 45
7, ND, 29
37, 30, 32
13, 33, 19
4.6 0.9
4.5
8.1
-5.7 (ND)
<-3.3 (99.8)
-5 (92.3)
-4.9 (75)
-4.1
1.1
10.4
>175
30.9
>175
114
1.7
5.7
1.1/9.4*
0.6/0.3**
2.6/3.1
19.0
8.6
5.1
3.3
1.0
>25
-4.9 (99.6)
2.1/3.3
20.8
Table 4. ADME profile of selected derivatives.1 Intrinsic Clearance (CLint) is in µL/min/mg in liver microsomes.2 IC50 data in
µM. Inhibition measured using midazolam as substrate.3 Data in µM measured from DMSO solution in buffer adjusted to pH6.8.4
Permeability is in cm/sec. 5 IC50 data in µM in dofetilide binding assay. * Efflux. ** Poor recovery (<30%).
Mouse PK
Rat PK
Parameter
CL1
Vss2
Cmax3 (PO)
AUC4
(PO)
%F
CL
Vss
Cmax
AUC
%F
11a
12a
12b
12c
20b
40
58
59
55
71
7.3
51
406
3
ND
15
ND
ND
291
ND
189
151
ND
ND
35
10.8
2.4
425
29
5980
123
59
2
13.7
ND
7187
ND
ND
19
ND
24
11.0
12.1
300
429
4959
4899
47
58
17.3
19.4
3268
3071
32
35
Table 5. Mouse and rat PK profile of compounds 11a, 12a-c and 20b. 1 mL/min/kg. 2 L/kg. 3 nM. 4 hrs*nM

16. Christensen, J. G.; Zou, H. Y.; Arango, M. E.; Li, Q.;
Lee, J. H.; McDonnell, S. R.; Yamazaki, S.; Alton, G. R.;
Mroczkowski, B.; Los, G. Mol. Cancer Therapeutics. 2007,
6(12), 3314.
Acknowledgments
17. Cui, J. J.; Tran-Dube, M.; Shen, H.; Nambu, M.; Kung,
P.-P.; Pairish, M.; Jia, L.; Meng, J.; Funk, L.; Botrous, I.;
McTigue, M.; Grodsky, N.; Ryan, K.; Padrique, E.; Alton, G.;
Timofeevski, S.; Yamazaki, S.; Li, Q.; Zou, H.; Christensen, J.;
Mroczkowski, B.; Bender, S.; Kania, R. S.; Edwards. M. P. J.
Med. Chem. 2011, 54(18), 6342.
The authors would like to thank Paul Calvin the compound
management group, John Isbell and the ADME and analytical
groups, Todd Groessl and the DMPK bioanalytical group, Muyun
Gao and Ken Ng and the Protein Sciences group, Frank Sun and
the Oncology Pharmacology group.
References and notes
18. Marsilje, T. H.; Pei, W.; Chen, B.; Lu, W.; Uno, T.; Jin,
Y.; Jiang, T.; Kim, S.; Li, N.; Warmuth, M.; Sarkisova, Y.; Sun,
F.; Steffy, A.; Pferdekamper, A. C.; Li, A. G.; Joseph, S. B.;
Kim, Y.; Liu , Tuntland, T.; Cui, X.; Gray, N. S.; Steensma, R.;
Wan, Y.; Jiang, J.; Chopiuk, G.; Li, J.; Gordon, W. P.;
Richmond, W.; Johnson, K.; Chang, J.; Groessl, T.; He, Y.-Q.;
Phimister, A.; Aycinena, A.; Lee, C. C.; Bursulaya, B.;
Karanewsky, D. S.; Seidel, H. M.; Harris, J. L. and Michellys, P.-
Y.; J. Med. Chem., 2013, 56 (14), pp 5675.
1. Morris, S. W.; Naeve, C.; Mathew, P.; James, P. L.;
Kirstein, M. N.; Cui, X.; Witte, D. P. ALK, the chromosome 2
gene locus altered by the t(2;5) in non-Hodgkin's lymphoma,
encodes a novel neural receptor tyrosine kinase that is highly
related to leukocyte tyrosine kinase (LTK). Oncogene. 1997, 14,
2175.
2. Chiarle, R.; Voena, C.; Ambrogio, C.; Piva, R.; Inghirami,
G. Nature Rev. Cancer. 2008, 8(1), 11.
19. Sakamoto, H.; Tsukaguchi, T.; Hiroshima, S.; Kodama,
T.; Kobayashi, T.; Fukami, T. A.; Oikawa, N.; Tsukuda, T.; Ishii,
N.; Aoki. Y. Cancer Cell. 2011, 19, 679.
3. Webb, T. R.; Slavish, J.; George, R. E.; Look, A. T.; Xue,
L.; Jiang, Q.; Cui, X.; Rentrop, W. B.; Morris, S. W. Expert Rev.
Anticancer Ther. 2009, 9(3), 331-.
.20. Kinoshita, K.; Kobayashi, T.; Asoh, K.; Furuichi, N.; Ito,
T.; Kawada, H.; Hara, S.; Ohwada, J.; Hattori, K.; Miyagi, T.;
Hong, W.-S.; Park, M.-J.; Takanashi, K.; Tsukaguchi, T.;
Sakamoto, H.; Tsukuda, T.; Oikawa, N. J. Med. Chem. 2011, 54,
6286.
4. Iwahara T.; Fujimoto J.; Wen D.; Cupples R.; Bucay N.;
Arakawa T.; Mori S.; Ratzkin B.; Yamamoto T. Oncogene.
1997, 14, 439.
5. Bilsland J. G.; Wheeldon A.; Mead A.; Znamenskiy P.;
Almond S.; Waters K. A.; Thakur M.; Beaumont V.; Bonnert T.
P.; Heavens R.; Whiting P.; McAllister G.; Munoz-Sanjuan I.
Neuropsychopharmacology. 2008, 33, 685.
21. Kuromitsu, S.; Mori, M.; Shimada, I.; Kondoh Y.;
Shindoh N.; Soga T.; Furutani T.; Konagai S.; Sakagami H.;
Nakata M.; Ueno Y.; Saito R.; Sasamata M.; Kudou M.
American Association for Cancer Research (AACR) Annual
Meeting in Orlando, Florida. 2011, abstract #2821.
6. Morris S. W.; Kirstein M. N.; Valentine M. B.; Dittmer K.
G.; Shapiro D. N.; Saltman D. L.; Look A. T. Science. 1994,
263, 1281.
.22. Lovly, C. M.; Heuckmann, J. M.; De Stanchina, E.; Chen,
H.; Thomas, R. K.; Liang, C.; Pao W. Cancer Res. 2011, 71(14),
4920.
7. Griffin, C. A.; Hawkins, A. L.; Dvorak, C.; Henkle, C.;
Ellingham,T.; Perlman, E. J. Cancer Res. 1999, 59, 2776.
8. Soda, M.; Choi, Y. L.; Enomoto, M.; Takada, S.;
Yamashita, Y.; Ishikawa, S.; Fujiwara, S.; Watanabe, H.;
Kurashina, K.; Hatanaka, H.; Bando, M.; Ohno, S.; Ishikawa, Y.;
Aburatani, H.; Niki, T.; Sohara, Y.; Sugiyama, Y.; Mano, H.
Nature. 2007, 448, 561.
23. Katayama, R.; Khan, T. M.; Benes, C.; Lifshits, E.; Eb, H.;
River, V. M.; Shakespeare, W. C.; Iafrate, A. J.; Jeffrey A.
Engelman; J. A., Shaw, A. T. Proceed. Nat. Acad. Sci. USA.
2011, 108(18), 7535.
24. Rivera, V. M.; Anjum, R.; Wang, F.; Zhang, S.; Keats, J.;
Ning, Y.; Wardwell, S. D.; Moran, L.; Ye, E.; Chun, D. Y.;
Mohemmad, K. Q.; Liu, S.; Huang, W.-S.; Wang, Y.; Thomas,
M.; Li, F.; Qi, J.; Miret, J.; Iuliucci, J. D.; Dalgarno, D.;
Narasimhan, N. I.; Clackson, T.; Shakespeare, W. C. American
Association for Cancer Research (AACR) Annual Meeting in
Washington, D.C., 2010, abstract #3623.
9. Mano, H. Cancer Sci. 2008, 99, 2349.
10. Shaw, A. T.; Solomon, B. Clin. Cancer Res. 2011, 17(8),
2081.
11. Osajima-Hakomori, Y.; Miyake, I.; Ohira, M.;
Nakagawara, A.; Nakagawa, A.; Sakai, R. Am. J. Pathol. 2005,
167, 213.
25. Johnson, T. W.; Richardson, P. F.; Bailey, S.; Brooun, A.;
Burke, B. J.; Collins, M. R.; Cui, J. J.; Deal, J. G.; Deng, Y.-L.;
Dinh, D.; Engstrom, L. D.; He, M., Hoffman, J.; Hoffman, R. L.;
Huang, Q.; Kania, R. S.; Kath, J. C.; Lam, H.; Lam, J. L.; Le, P.
T.; Lingardo, L.; Liu, W.; McTigue, M.; Palmer, C. L.; Sach, N.
W.; Smeal, T.; Smith, G. L.; Stewart, A. E.; Timofeevski, S.;
Zhu, H.; Zhu, J.; Zou, H. Y.; Edwards, M. P.; J. Med. Chem.,
2014, 57 (11), 4720.
12. Mosse, Y. P.; Laudenslager, M.; Longo, L.; Cole, K.A.;
Wood, A.; Attiyeh, E. F.; Laquaglia,M. J.; Sennett, R.; Lynch, J.
E.; Perri, P.; Laureys, G.; Speleman, F.; Kim, C.; Hou, C.;
Hakonarson, H.; Torkamani, A.; Schork, N. J.; Brodeur, G. M.;
Tonini, G. P.; Rappaport, E.; Devoto, M.; Maris., J. M. Nature.
2008, 455, 930.
13. Azarova, A. M.; Gautam, G.; George, R. E. Seminars in
Cancer Bio. 2011, 21, 267.
26. Friboulet, L., Li, N., Katayama, R., Lee, C., Gainor, J. F.,
Crystal, A. S., Michellys, P.-Y., Awad, M. M., Yanagitani, N.,
Kim, S., Pferdekamper A. C., Li, J., Kasibhatla, S., Sun, F., Sun.,
X., Hua, S., McNamara, P., Mahmood, S., Lockerman, E. L.,
Fujita, N., Nishio, M., Harris, J. L., Shaw, A. T., Engelman, J. A.,
Can. Disc. 2014, 10, 662.
14. Tuma, R. S. J. Nat. Cancer Inst. 2012, 104(2), 87-88.
15. Ren, H.; Tan, X. Z.; Crosby, C.; Haack, H.; Ren, J.-M.;
Beausoleil, S.; Moritz, A.; Innocenti, G.; Rush, J.; Zhang, Y.;
Zhou, X.-M.; Gu, T.-L.; Ynag, Y.-F.; Comb, M. J. Cancer Res.
2012, 72, 3312.

27. PDB code 4MKC.
28. Chen, B.; Jiang, T.; Marsilje, T. H.; Michellys, P.-Y.;
Nguyen, T. N.; Pei, W.; Wu, B.; Gao, Z.; Ge, Y.; Huang, C.; Li,
Y.; WO 2009126515 A1
29. PDB code 4Z55.
30. No attempts were done to try to improve activity using
these single bicyclic hinge moieties; since both 24 and 25 possess
a hinge binding moiety, it may be possible to re-optimize the
compounds using divergent SAR directions.
31. The study was intended to be conducted for 2 weeks.
However, due to the very aggressive growth rate of these tumors,
the vehicle animals were sacrificed after 9 days for humane
reasons.