Novel bis-ortho-alkoxy-para-piperazinesubstituted-2,4-dianilinopyrimidines (KRCA-0008) as potent and selective ALK inhibitors for anticancer treatment

Article abstract of DOI:10.1016/j.bmcl.2013.08.090

The synthesis of bis-ortho-alkoxy-para-piperazinesubstituted-2,4- dianilinopyrimidines is described and their structure-activity-relationship to anaplastic lymphoma kinase (ALK) is presented. KRCA-0008 is selective and potent to ALK and Ack1, and displays drug-like properties without hERG liability. KRCA-0008 demonstrates in vivo efficacy comparable to Crizotinib in xenograft mice model.

Full text of DOI:10.1016/j.bmcl.2013.08.090

Bioorganic & Medicinal Chemistry Letters 23 (2013) 6192–6196
Contents lists available at ScienceDirect
Bioorganic & Medicinal Chemistry Letters
journal homepage: www.elsevier.com/locate/bmcl
Novel bis-ortho-alkoxy-para-piperazinesubstituted-2,
4-dianilinopyrimidines (KRCA-0008) as potent and selective
ALK inhibitors for anticancer treatment
Chi Hoon Park a, Hyeonjeong Choe a,b, In-Young Jang a, So Yeong Kwon a, Muhammad Latif a,b,c
,
Heung Kyoung Lee a, Hyeon Ji Lee a,d, Eun Hye Yang a, Jeong In Yun a,e, Chong Hak Chae a,c, Sung Yun Cho a,
Sang Un Choi a, Jae Du Ha a, Heejung Jung a, Hyoung Rae Kim a, Pilho Kim a, Chong Ock Lee a, Chang-Soo Yun a,
Kwangho Lee a,b,
⇑
a Bio-Organic Science Division, Korea Research Institute of Chemical Technology, PO Box 107, Daejeon 305-600, Republic of Korea
b Medicinal & Pharmaceutical Chemistry, University of Science & Technology, Daejeon 305-350, Republic of Korea
c Korea Chemical Bank, Daejeon 305-600, Republic of Korea
d College of Pharmacy, Chungbuk National University, Cheongju 361-763, Republic of Korea
e College of Pharmacy, Chungnam National University, Daejeon 305-764, Republic of Korea
a r t i c l e i n f o
a b s t r a c t
Article history:
The synthesis of bis-ortho-alkoxy-para-piperazinesubstituted-2,4-dianilinopyrimidines is described and
their structure–activity-relationship to anaplastic lymphoma kinase (ALK) is presented. KRCA-0008 is
selective and potent to ALK and Ack1, and displays drug-like properties without hERG liability. KRCA-
0008 demonstrates in vivo efficacy comparable to Crizotinib in xenograft mice model.
Ó 2013 Elsevier Ltd. All rights reserved.
Received 24 July 2013
Revised 19 August 2013
Accepted 23 August 2013
Available online 4 September 2013
Keywords:
ALK
ACK1
Kinases
NSCLC
Diaminopyrimidine
Inhibitors
N-Acetylpiperidine
Anaplastic lymphoma kinase (ALK) belongs to the insulin recep-
tor superfamily of tyrosine kinase. ALK-fused genes such as NPM-
ALK, EML4-ALK, KIF5-ALK are putative oncogenes and responsible
for many cancers including anaplastic large-cell lymphomas
(ALCL), inflammatory myofibroblastic tumors (IMT), diffuse large
B cell lymphoma (DLBCL) and a variety of solid tumor types.1
Crizotinib (XalkoriÒ, Pfizer) is the first EML4-ALK inhibitor in
clinical use for non-small-cell-lung cancer (NSCLC) treatment.
However, despite dramatic responses to Crizotinib, its efficacy is
ultimately limited by the development of acquired drug resistance
that occurred within a year including EML4-ALK L1196M, F1174L,
R1275Q and G1269S and grade 3 or 4 adverse event of ALT and AST
elevation (ꢀ10%).2
dine ALK inhibitors, NVP-TAE684 and AP26114, reportedly retain
potency over Crizotinib-resistant ALK mutants.3 They possess
either a sulfone or phosphine oxide at the aniline ortho position
of the C4 pyrimidine to make a selective hydrogen interaction to
ALK Lys1150. Of many ALK pyrimidine analogs, Cephalon recently
reported a methoxyl instead of the ortho carbonyl and achieved
moderate potency but excellent selectivity over homologous insulin
receptor (IR) which is likely responsible for compound-dependent
glucose fluctuation in preclinical animal study.4 Inspired by this
report, we decided to explore the alkoxy structure–activity-
relationship (SAR) and improve its potency by replacing the mor-
pholines with more polar piperazine derivatives which in general
allows lower plasma–protein-binding (PPB) for improved efficacy.
A general synthetic chemistry is outlined in Scheme 1. 5-Fluoro-
2-nitrophenol 1 was alkylated with various alkyl halide in the pres-
ence of potassium carbonate in N,N-dimethylformamide. Thus
alkylated arylfluoride 2 was then reacted with N-acetylpiperazine
to provide 1-(4-(3-alkoxy-4-nitrophenyl)piperazin-1-yl)ethanone
3. Subsequent hydrogenation afforded the key building block
To overcome these limitations, other novel ALK inhibitors such
as CH5424802, AP26113, NVP-TAE684, and LDK378 have been
studied and are currently under development (Fig. 1). The pyrimi-
⇑
Corresponding author. +82 42 860 7176
E-mail address: kwangho@krict.re.kr (K. Lee).
0960-894X/$ - see front matter Ó 2013 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.bmcl.2013.08.090

C. H. Park et al. / Bioorg. Med. Chem. Lett. 23 (2013) 6192–6196
6193
NH
O
S
P
N
N
N
O
O
N
N
NH
NH
Cl
N
Cl
F
Cl
N
1
N
N
N
N
O
N
N
H
N
N
H
NH2
O
Cl
O
Crizotinib
AP26113
NVP-TAE684
O
O
O
S
O
N
O
O
N
N
H
N
NH
N
NH
NH
N
Cl
Cl
N
NC
N
N
H
N
N
H
O
O
O
CH5424802
LDK378
Cephalon
Figure 1. ALK Inhibitors.
O
N
F
OH
F
O
a
b
c
R
N
O
R
NO2
NO2
2
1
3
NO2
O
O
N
N
N
O
R
O
N
O
O
R
N
N
e
d
N
NH
N
NH
O
N
R
Cl
Cl
N
NH2
N
N
H
N
Cl
6
4
5
O
Scheme 1. Reagents and conditons: (a) R-I, K2CO3, DMF, 50 °C, 3 h (83–96%); (b) N-acetylpiperazine, K2CO3, DMF, 80 °C, overnight (79–93%); (c) H2, Pd/c, EtOH overnight
(quantitative); (d) 2,4,5-trichloropyrimidine, K2CO3, DMF, 80 °C, overnight (72-90%); (e) 1-(4-(4-amino-3-mathoxyphenyl)piperazin-1-yl)ethanone, HCl, ethoxyethanol,
115 °C, 3 h (35–83%).
Table 1
1-(4-(4-amino-3-alkoxyphenyl)piperazin-1-yl)ethanone 4 as a dark
Comparison of N-acetylpiperazine and morpholine on the A ring
solid. The aniline 4 was then condensed with 2,4,5-trichloropyrim-
idine in N,N-dimethylformamide at 80 °C to make a regioselective
adduct 5. The adduct
5 was heated with 1-(4-(4-amino-3-
X
methoxyphenyl)piperazin-1-yl)ethanone in the presence of cata-
lytic amounts of hydrogen chloride in ethoxyethanol at 115 °C to
provide an ALK inhibitor 6.5
N
O
O
NH
N
Cl
N
First of all, we compared the N-acetylpiperazine substitution on
the A ring (KRCA-0008) to the morpholine analogue compound
7on ALK activity (Table 1). To our surprise, compound 7 was unex-
pectedly less potent and the N-acetylpiperazine analogue (KRCA-
0008) was far superior to compound 7 in both biochemical and
cellular assay.
N
N
N
H
O
Compound
X
ALK wt.IC50 (nM) H1993 IC50
(
l
M) H3122 IC50 (lM)
Although we do not fully understand the role of N-acetylpiper-
azine on the A ring at this time, we next explored the ortho-meth-
oxy effect on both A & C rings as shown in Table 2. When compared
to compound 8 (no o-OMe on A & C rings), compound 9 (o-OMe
on the C ring) displays slightly improved biochemical ALK wt.
activity and similar H3122 cell (EML4-ALK driven) proliferation po-
tency with slightly improved H1993 cell (c-MET driven) selectivity.
When the o-OMe is placed on the A ring (compound 10), ALK po-
tency in both biochemical and H3122 cell improved noticeably,
KRCA-0008 NAc
12
2600
3.6
4.5
0.08
3.3
7
O
although the H1993 selectivity decreased from that of compound
8. Combined with these results, KRCA-0008, a bis-ortho-OMe

6194
C. H. Park et al. / Bioorg. Med. Chem. Lett. 23 (2013) 6192–6196
Table 2
The ortho-methoxy effect
O
The A ring o-Methoxy brings ALK activity in both biochemical & cellular
assay while keeping IR selectivity.
N
A
N
R1
NH
O
N
C
N
Cl
N
N
N
H
B
R2
The C ring o-Methoxy brings ALK activity and
selectivity over other kinases.
Compound
R1
R2
ALK wt. IC50 (nM)
ALK L1196 M IC50 (nM)
IR IC50 (nM)
H1993 IC50
(
lM)
H3122 IC50 (lM)
Crizotinib
8
9
10
36
300
64
18
12
220
naa
330
210
75
350
na
300
660
210
0.083
2.1
9.1
0.98
3.6
0.28
0.91
0.81
0.096
0.08
H
H
OMe
OMe
H
OMe
H
KRCA-0008
OMe
a
na: not available.
analog attached on both the A and C rings, gratifyingly improved
the biochemical activity and H3122 cellular potency significantly
while retained good H1993 selectivity.6 This observation was in
general consistent with other close analogs (data not shown) and
we could draw a conclusion regarding the ortho-OMe effect on
the pyrimidine ALK inhibitors: The A ring o-Me b rings ALK activity
in both biochemical & cellular assay while keeping IR selectivity,
and the C ring o-Me adds ALK activity and selectivity over other
kinases.
Encouraged by this observation, a systematic SAR study on the
A ring ortho-alkoxy was conducted as summarized in Table 3. The
A ring methoxy-containing KRCA-0008 was proven an optimal
substituent in terms of potency after all and its bigger size deriva-
tives were detrimental in both biochemical and H3122 cellular
activities (compounds 11–15). Since difluoromethoxy analog
(compound 16) is equipotent with KRCA-0008, the alkoxy size
on the ortho position proved critical on their ALK activity. It is note-
worthy that the preparation of the hydroxyl analog (–OH) on the
ortho position failed presumably because of the instability of
the 2-hydroxy-1,4-diaminobenzene moiety.7 Besides alkoxys on
the A ring ortho position, halide and amino replacements were also
examined. Chlorine and fluorine derivatives (compounds 17, 18)
Table 3
The Aring ortho-substitution SAR
O
N
N
R
O
NH
N
N
Cl
N
N
N
H
O
Compound
R
ALK wt. ALK
IR
H1993 H3122 BaF3 EML4-
IC50 IC50 ALK L1196 M
M) M) IC50 M)
IC50
L1196 M IC50
IC50 (nM) (nM)
(nM)
(l
(l
(l
Crizotinib
9
KRCA-0008 OMe 12
36
64
220
330
75
230
360
360
na
350
300
210
>1000 6.9
>1000 7.3
>1000 5.9
na
na
300
na
na
na
na
0.083 0.28
0.88
naa
0.068
0.27
na
na
na
2
H
9.1
3.6
0.81
0.08
0.96
0.84
0.41
1.4
11
12
13
14
15
16
17
18
19
20
OEt
25
On-Pr 38
Oi-Pr 33
1000
>1000 na
OCHF2 10
OBn
OPh
9.7
na
2.1
na
na
na
na
4.1
41
44
280
na
0.024 0.15
0.19 0.17
0.091 0.44
Cl
F
7
11
Nt-Bu 580
NHPh >1000 na
4.4
>10
na
na
a
na: not available.
Figure 2. Immunoblot analysis of selected compounds.

C. H. Park et al. / Bioorg. Med. Chem. Lett. 23 (2013) 6192–6196
6195
Table 4
exposure (Cbrain/Cplasma = ꢀ0.02). It has good liver microsomal sta-
bility (% remaining after 30 min: 52% in mouse, 89% in rat, 72% in
human) and little to no CYP inhibition (1A2, 2C9, 2D6, 3A4 @
Pharmacokinetic parameters of KRCA-0008
ICR micea
Sprague Dawley ratsa
10
lM). It does not appear to cause hERG blockade (patch clamp
Parameters
IV, 5 mg/kgb PO, 5 mg/kgb IV, 5 mg/kgc PO, 20 mg/kgc
IC50 = 30
l
M) and is negative on Ames test (1000 lg/plate), chro-
Tmax (h)
—
—
0.5
0.54 0.174
—
—
1.33 0.578
2.51 0.098
mosomal aberration assay and micronucleus assay.10
Cmax
(
lg/mL)
KRCA-0008 also shows promising pharmacokinetic parameters
in both mice and rat (oral bioavailability = 66–94.5%) as summa-
rized in Table 4.11 Finally, KRCA-0008 shows a modest tumor
growth inhibition in vivo activity in H3122 human lung cancer
bearing mice model comparable to Crizotinib without significant
body weight change (Fig. 3).12 It is important to mention the
KRCA-0008 25 mpk and 50 mpk groups did not show dose-depen-
dent tumor growth inhibition.
T1/2 (h)
AUCt (
AUC1 (
1.31 0.381 2.23 1.15
1.68 0.23 1.59 0.226
g h/mL) 1.72 0.278 1.62 0.23
2.72 0.903 2.19 0.579
4.95 1.15 13.1 3.49
4.95 1.15 13.1 3.52
l
l
CL (L/kg/h)
Vss (L/kg)
F (%)
g th/mL)
2.96 0.438
4.352 1.12
—
—
—
1.04 0.216
1.504 0.154
—
—
—
94.5
66.2
a
b
c
Values are means of data from three animals.
Vehicle: DMSO 5%, PEG400 50%, DW 45% (2.5 mg/mL).
Vehicle: DMSO 5%, PEG400 40%, DW 55% (2.5 mg/mL).
In summary, the synthesis of bis-ortho-alkoxy-para-piperazine-
substituted-2,4-dianilinopyrimidines is described and their struc-
ture-activity-relationship study to an aplastic lymphoma kinase
(ALK) is discussed. KRCA-0008 is selective and potent to ALK and
Ack1, and has drug-like properties without hERG concerns.
KRCA-0008 demonstrates in vivo efficacy comparable to Crizotinib
in xenograft mice model. Currently KRCA-0008 analogs are under
investigation and those will be reported in the future.
possess similar biochemical and cellular potency to KRCA-0008,
and amine analogs (compound 19, 20) resulted in a significant loss
in the biochemical ALK wt. activities.
KRCA-0008 is potent to ALK wt. and mutants in biochemical as-
say (ALK wt. IC50 = 12 nM, ALK L1196 M IC50 = 75 nM, ALK C1156Y
IC50 = 4 nM, ALK F1174L IC50 = 17 nM, ALK R1275Q IC50 = 17 nM). It
has 0.08
liferation and possesses 3.6
l
M IC50 in H3122 (EML4-ALK driven tumor cell) cell pro-
M cellular selectivity to H1993 (c-
Acknowledgment
l
MET driven non small cell lung cancer). Immunoblotting of phos-
pho-ALK and its downstream targets (phospho-Akt and phospho-
Erk) was carried out in H3122 (EML4-ALK) and A549 (ALK wt.) cells
and proved specific inhibition to EML4-ALK driven H3122 cell
using compound 11 and KRCA-0008as depicted in Figure 2. Ambit
assay (>100 kinase panel assay done by Millipore service) shows
compound 11 is very potent only to ALK (IC50 = 25nM) and ACK1
(IC50 = 18nM), and very selective over the rest other kinases includ-
This work was supported by the Korea Research Institute of
Chemical Technology (KK-1303), the Korea Chemical Bank (SI-
1305) and Ministry of Science, ICT and Future Planning of Korea
(KN-1235), and authors thank Sanghun Alex Lee for manuscript
proofreading.
Supplementary data
ing IGF-1R and IR (insulin receptor) at 1
KRCA-0008 retains good drug-like properties: good water-solu-
bility (54 M in 5% DMSO–water, 150 M in 5% DMSO–PBS buffer)
with moderate plasma protein binding (93% in rat) and low brain
l
M.8,9
Supplementary data associated with this article can be found,
in the online version, at http://dx.doi.org/10.1016/j.bmcl.2013.
08.090.
l
l
Figure 3. H3122 xenograft in mice12
.

6196
C. H. Park et al. / Bioorg. Med. Chem. Lett. 23 (2013) 6192–6196
J = 8.5 Hz, 1 H), 8.19 (d, J = 8.5 Hz, 1 H), 8.03 (s, 1 H), 7.74 (s, 1 H), 7.27 (s, 1 H),
References and notes
6.57–6.52 (m, 4 H), 4.14 (q, J = 6.7 Hz, 2 H), 3.90 (s, 3 H), 3.82–3.78 (m, 4 H),
3.68–3.65 (m, 4 H), 3.18–3.10 (m, 8 H), 2.19 (s, 3 H), 2.18 (s, 3 H), 1.51 (t,
J = 6.7 Hz, 3 H); Mass (M+H+) calcd for C31H39ClN8O4 622.27, found 623.01;
1. (a) Koivunen, J. P.; Mermel, C.; Zejnullahu, K.; Murphy, C.; Lifshits, E.; Holmes,
A. J.; Choi, H. G.; Kim, J.; Chiang, D.; Thomas, R.; Lee, J.; Richards, W. G.;
Sugarbaker, D. J.; Ducko, C.; Lindeman, N.; Marcoux, J. P.; Engelman, J. A.; Gray,
N. S.; Lee, C.; Meyerson, M.; Jänne, P. A. Clin. Cancer Res. 2008, 14(13), 4275; (b)
Sasaki, T.; Rodig, S. J.; Chirieac, L. R.; Jänne, P. A. Eur. J. Cancer 2010, 46, 1773; (c)
Galkin, A. V.; Melnick, J. S.; Kim, S.; Hood, T. L.; Li, N.; Li, L.; Xia, G.; Steensma, R.;
Chopiuk, G.; Jiang, J.; Wan, Y.; Ding, P.; Liu, Y.; Sun, F.; Schultz, P. G.; Gray, N. S.;
Warmuth, M. Proc. Natl. Acad. Sci. U.S.A. 2007, 270; Minoo, P.; Wang, H.-Y. Int. J
Clin. Exp. Pathol. 2012, 5(5), 397.
2. (a) Cui, J. J.; Tran-Dubé, M.; Shen, H.; Nambu, M.; Kung, 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, 6342; (b) 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. (a) Choi, Y. L.; Soda, M.; Yamashita, Y.; Ueno, T.; Takashima, J.; Nakajima, T.;
Yatabe, Y.; Takeuchi, K.; Hamada, T.; Haruta, H.; Ishikawa, Y.; Kimura, H.;
Mitsudomi, T.; Tanio, Y.; Mano, H. N. Eng. J. Med. 2010, 363(18), 1734; (b)
Katayama, R.; Khan, T. M.; Benes, C.; Lifshits, E.; Ebi, H.; Rivera, V. M.;
Shakespeare, W. C.; Iafrate, A. J.; Engelman, J. A.; Shaw, A. T. Proc. Natl. Acad. Sci.
USA 2011, 108, 7535; (c) Sasaki, T.; Koivunen, J.; Ogino, A.; Yanagita, M.;
Nikiforow, S.; Zheng, W.; Lathan, C.; Marcoux, J. P.; Du, J.; Okuda, K.; Capelletti,
M.; Shimamura, T.; Ercan, D.; Stumpfova, M.; Xiao, Y.; Weremowicz, S.;
Butaney, M.; Heon, S.; Wilner, K.; Christensen, J. G.; Eck, M. J.; Wong, K.-K.;
Lindeman, N.; Gray, N. S.; Rodig, S. J.; Jänne, P. A. Cancer Res. 2011, 71, 6051; (d)
Zhang, S.; Wang, F.; Keats, J.; Zhu, X.; Ning, Y.; Wardwell, S. D.; Moran, L.;
Mohemmad, Q. K.; Anjum, R.; Wang, Y.; Narasimhan, N. I.; Dalgarno, D.;
Shakespeare, W. C.; Miret, J. J.; Clackson, T.; Rivera, V. M. Chem. Biol. Drug Des.
2011, 78, 999.
Analytics
for
KRCA-0008:
1,10-(4,40-(((5-chloropyrimidine-2,4-
diyl)bis(azanediyl))bis(3-methoxy-4,1-phenylene))bis(piperazine-4,1-
diyl))diethanone: 1H NMR (300 MHz, CDCl3) d 8.29 (d, J = 8.6 Hz, 1 H), 8.15 (d,
J = 8.6 Hz, 1 H), 8.01 (s, 1 H), 7.57 (s, 1 H), 6.47–6.57 (m, 4 H), 3.92 (s, 3 H), 3.88
(s, 3 H), 3.77–3.82 (m, 4 H), 3.63-3.69 (m, 4 H), 3.06–3.18 (m, 8 H), 2.16 (s, 3 H),
2.15 (s, 3 H); Mass (M + H+) calcd for C30H37ClN8O4 608.26, found 609.17.
7. Various attempts to prepare the ortho-hydroxyl analog has been made but
resulted in intractable products.
8. Activated Cdc42-associated kinase (Ack1) is non-receptor tyrosine kinase and a
critical early transducer of variety of extracellular growth factor stimuli
including heregulin, insulin, EGF and PDGF signaling. For more detail, see: (a)
Mahajan, N. P.; Whang, Y. E.; Mohler, J. L.; Earp, H. S. Cancer Res. 2005, 65(22),
10514; (b) Mahajan, K.; Coppola, D.; Challa, S.; Fang, B.; Chen, Y. A.; Zhu, W.;
Lopez, A. S.; Koomen, J.; Engelman, R. W.; Rivera, C.; Muraoka-Cook, R. S.;
Cheng, J. Q.; Schönbrunn, E.; Sebti, S. M.; Earp, H. S.; Mahajan, N. P. PLos ONE
2010, 5(3), e9646; (c) Miduturu, C. V.; Deng, X.; Kwiatkowski, N.; Yang, W.;
Brault, L.; Filippakopoulos, P.; Chung, E.; Yang, Q.; Schwaller, J.; Knapp, S.; King,
R. W.; Lee, J.-D.; Herrgard, S.; Zarrinkar, P.; Gray, N. S. Chem. Biol. 2011, 18, 868.
9. Ambit assay results (@ 1 lM of compounds 9, 11) and selected IC50s are
attached to Supplementary data
10. Flamand, N.; Meunier, J.-R.; Meunier, P.-A.; Agapakis-Caussé, C. Toxicol. In Vitro
2001, 15, 105; (b) Fenech, M. Mutation Res. 2000, 455, 81; Jena, G. B.; Kaul, C. L.;
Ramarao, P. J. Indian Pharmacol. 2002, 34, 86.
11. There is a concern of reactive metabolite formation (GSH adduct) on the 1,4-
diaminobenzene moiety resulting in fast elimination in bile. However, KRCA-
0008 shows moderate clearance in both mice and rat. For more information,
see: (a) Wells-Knecht, K. J.; Ott, G. R.; Cheng, M.; Wells, G. J.; Breslin, H. J.;
Gingrich, D. E.; Weinberg, L.; Mesaros, E. F.; Huang, Z.; Yazdanian, M.; Ator, M.
A.; Aimone, L. D.; Zeigler, K.; Dorsey, B. D. Chem. Res. Toxicol. 1994, 24, 2011; (b)
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, B.; 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.; Michellys, P.-Y. J. Med. Chem.
2013, 56, 5675.
12. Effects of compounds on growth of H3122 human lung cancer in nude mice. H3122
cells were implanted S.C. into the right flanks of female nude mice. Drug
treatment was initiated after tumor volumes reached about 200 mm3. The data
are expressed as the mean tumor size S.E. 8 mice/group, Formulation: 20%
PEG400, 3% Tween80 in DDW, Route: PO, Schedule: 14 days (5 days on, 2 days
off) [KRCA-0008: 25 mpk & 50 mpk, p.o., BID; Crizotinib: 25 mpk, p.o., QD].
4. Mesaros, E. F.; Burke, J. P.; Parrish, J. D.; Dugan, B. J.; Anzalone, A. V.; Angeles, T.
S.; Albom, M. S.; Aimone, L. D.; Quail, M. R.; Wan, W.; Lu, L.; Huang, Z.; Ator, M.
A.; Ruggeri, B. A.; Cheng, M.; Ott, G. R.; Dorsey, B. D. Bioorg. Med. Chem. Lett.
2011, 21, 463.
5. (a) Ott, G. R.; Tripathy, R.; Cheng, M.; McHugh, R.; Anzalone, A. V.; Underiner, T.
L.; Curry, M. A.; Quail, M. R.; Lu, L.; Wan, W.; Angeles, T. S.; Albom, M. S.;
Aimone, L. D.; Ator, M. A.; Ruggeri, B. A.; Dorsey, B. D. ACS Med. Chem. Lett. 2010,
1, 493; (b) Gingrich, D. E.; Lisko, J. G.; Curry, M. A.; Cheng, M.; Quail, M.; Lu, L.;
Wan, W.; Albom, M. S.; Angeles, T. S.; Aimone, L. D.; Haltiwanger, R. C.; Wells-
Knecht, K.; Ott, G. R.; Ghose, A. K.; Ator, M. A.; Ruggeri, B.; Dorsey, B. D. J. Med.
Chem. 2012, 55, 4580.
6. Analytics for compound 11: 1-(4-(4-((4-((4-(4-acetylpiperazin-1-yl)-2-
ethoxyphenyl)amino)-5-chloropyrimidin-2-yl)amino)-3-
methoxyphenyl)piperazin-1-yl)ethanone: 1H NMR (300 MHz, CDCl3) d 8.36 (d,