In vivo and in vitro SAR of tetracyclic MAPKAP-K2 (MK2) inhibitors. Part i

Article abstract of DOI:10.1016/j.bmcl.2010.04.024

Pyrrolo[2,3-f]isoquinoline based amino acids, tetracyclic lactams and cyclic ketone analogues are described as novel MK2 inhibitors with IC ₅₀ as low as 5 nM and good selectivity profiles against a number of related kinases including ERK, p38α and JNKs. TNFα release was suppressed from human peripheral blood mononuclear cells (hPBMCs), and a representative compound inhibited LPS induced TNFα release in mice illustrating the potential of this series to provide orally active MK2 inhibitors.

Full text of DOI:10.1016/j.bmcl.2010.04.024

Bioorganic & Medicinal Chemistry Letters 20 (2010) 4715–4718
Contents lists available at ScienceDirect
Bioorganic & Medicinal Chemistry Letters
journal homepage: www.elsevier.com/locate/bmcl
In vivo and in vitro SAR of tetracyclic MAPKAP-K2 (MK2) inhibitors. Part I
*
Laszlo Revesz , Achim Schlapbach, Reiner Aichholz, Roland Feifel, Stuart Hawtin, Richard Heng,
Peter Hiestand, Wolfgang Jahnke, Guido Koch, Markus Kroemer, Henrik Möbitz, Clemens Scheufler,
Juraj Velcicky, Christine Huppertz
Novartis Institutes for BioMedical Research, CH-4002 Basel, Switzerland
a r t i c l e i n f o
a b s t r a c t
Article history:
Pyrrolo[2,3-f]isoquinoline based amino acids, tetracyclic lactams and cyclic ketone analogues are
Received 11 March 2010
Revised 7 April 2010
Accepted 7 April 2010
Available online 13 April 2010
described as novel MK2 inhibitors with IC50 as low as 5 nM and good selectivity profiles against a number
of related kinases including ERK, p38a and JNKs. TNFa release was suppressed from human peripheral
blood mononuclear cells (hPBMCs), and a representative compound inhibited LPS induced TNF
a release
in mice illustrating the potential of this series to provide orally active MK2 inhibitors.
Ó 2010 Elsevier Ltd. All rights reserved.
Keywords:
MAPKAP-K2
MK2
TNFa
Inhibition
Antiinflammatory
In vivo
Rheumatoid arthritis
Mitogen-activated protein kinases (MAPKs) belong to the Ser/
Thr kinase family1, control cytoskeletal architecture, cell-cycle
progression and are implicated in inflammation and cancer.2
The p38/MAPKAP kinase-2 (MAPKAP-K2; MK2) cascade plays a
pivotal role in the production of proinflammatory cytokines,
Bromide 4—the precursor of MK2 inhibitors 5–8—was prepared
from 3-chloro-5-aminoisoquinoline (Scheme 1), which was con-
densed with 3-oxobutyric acid tert-butyl ester to an enamine that
cyclised under oxidative Pd(OAc)2/Cu(OAc)2 catalysis9 to provide
pyrroloisoquinoline 3. BOC-protection followed by NBS treatment
gave 4, which was reacted with methylamine, ammonia, meth-
ylhydrazine and tert-butyl N-hydroxycarbamate. Suzuki10 coupling
and deprotection yielded amino acids 5A, 6A and the amino acid
precursor of 7A, which was cyclised with EDCI. Suzuki coupling
with 8 (R1 = Cl) was successful and yielded 8E, while 7 (R1 = Cl)
did not survive the coupling conditions.
such as TNFa . Moreover, MK2 knock-out mice
, IL-6 and IFNc 3
are resistant to developing disease in arthritis models.4 Thus,
MK2 has emerged as a highly desirable target in the search for
efficacious and safe anti-inflammatory drugs. Several reports on
MK2 inhibitors from a variety of structural classes were pub-
lished5 including pyrrolopyrimidinones6 and aminopyrazoles.7
Recently the difficulties in developing MK2 inhibitors have been
reviewed.8
Boronic acids or their pinacol esters employed to introduce side
chains R1 = A–C and E–K were either commercially available or
prepared as described previously.11
Our search for MK2 inhibitors began with high throughput
screening that identified the pyrrolo[2,3-f]isoquinoline amide 1
Amino acid 5A was a potent and rather selective MK2 inhibitor
as a moderate MK2 inhibitor with an IC50 value of 3.8
l
M
(IC50 = 0.014
0.735 M). Potential prodrugs of 5A, such as the methyl ester
and the primary amide (not shown in Table 1) were considerably
weaker (ester: 11 M; amide: 1.2 M). The high potency of 5A
may be explained by possible salt formation with Lys93 and
Asp207 of MK2 (see X-ray analysis, Figure 2). The N-methyl ana-
logue 6A led to a drop in MK2 inhibitory potency and cellular activ-
ity. Lactams 7A and 8E were both potent MK2 inhibitors with 7A
lM; Table 1), showing cellular efficacy (IC50 =
( Fig. 1). 1 demonstrated structural similarities to the recently
disclosed MK2 inhibitors 25f with a pyrrolopyridine scaffold and
submicromolar IC50. A chemistry program was initiated aimed at
enhancing potency of MK2 inhibitors by combining structural fea-
tures of 1 and 2. Described herein are the resulting novel MK2
inhibitors 5–8, 10, 11, 13, 14 and their SAR.
l
l
l
also inhibiting LPS induced TNF
(IC50 = 0.046 M). The latter effect may partly be due to off-target
kinase inhibition, since 7A inhibited 6 out of 26 kinases12 with
a release from hPBMCs
l
* Corresponding author. Fax: +41 61 324 3273.
E-mail address: laszlo.revesz@pharma.novartis.com (L. Revesz).
0960-894X/$ - see front matter Ó 2010 Elsevier Ltd. All rights reserved.
doi:10.1016/j.bmcl.2010.04.024

4716
L. Revesz et al. / Bioorg. Med. Chem. Lett. 20 (2010) 4715–4718
R1:
N
N
O
O
R
NH2
N
H
N
H
NH
HN
1
2
Figure 1. MK2 inhibitors. Compound 1: HTS-hit. Compound 2: Pfizer’s MK2
N
N
O
O
O
inhibitors.5f
F
R
C
A
B
D
I
J
K
N
N
E (R=4-OMe)
F (R=3-F-4-OMe)
G (R=2-F)
N
O
O
O
N
O
O
IC50 <1
of TNF
l
M. Since JNK2-inhibition may contribute to the inhibition
a
13 release and toxicity, MK2 inhibitors with minimal JNK2-
H (R=2-CF3 )
inhibition are desirable. 8E showed a 23.7-fold preference for MK2
over JNK2 (Table 1), inhibited hsp27 phosphorylation with
IC50 = 0.87 lM and LPS induced TNFa release with IC50 = 2.36 lM,
Scheme 1b. R1-substituents.
which supports the notion, that its cellular efficacy was MK2-
driven.
Table 1
Tetracyclic MK2 inhibitors 10 and 11 with a 5-membered lac-
tam-ring were prepared according to Scheme 2 from 4, which
was reacted with MeNH2 or NH3. The resulting amines were depro-
tected to amino acids and then cyclised to 9 with EDCI. Suzuki cou-
pling with R1–B(OH)2 or its pinacol ester provided 10A–C, 10E–K
and 11A. 3-Fluoroaniline and 9 delivered 10D via Buchwald14 reac-
tion conditions. Styrene derivatives 10A–C (Table 2) were potent
Compd MK2a
M)
TNFb
M)
p-hsp27c
M)
JNK2d
M)
Selectivityd
(
l
(
l
(
l
(
l
5A
6A
7A
8E
0.014
0.060
0.025
0.023
0.735
1.240
0.046
2.359
nt
nt
nt
nt
nt
nt
2
nt
6
0.875
0.547
2
IC50 values are reported as the mean of P2 experiments with a standard deviation
MK2 inhibitors (IC50 60.010
(IC50 60.034 M) by 10A–C possibly contributed to the potent
inhibition of LPS induced TNF in hPBMCs (IC50 60.014 M). Ani-
line derivative 10D had 75-fold higher affinity for JNK2
(IC50 = 0.001 M) than for MK2. Compound 10H among the phenyl
derivatives 10E–I demonstrates the importance of coplanarity be-
tween the tetracyclic core and the R1-substituent. The large 2-tri-
fluoromethyl group in 10H forces the phenyl group out of plane
lM), but highly unselective. JNK2
of less than 50%.
a
MK2 enzyme assay is performed as described.7
l
b
Inhibition of LPS stimulated release of TNF
a
from hPBMC is performed as
a
l
described.7
a
hsp27 phosphorylation is performed as described.7
c
d
Number of kinases inhibited with IC50 <1 l
M out of a panel of 26 kinases.12 nt:
l
not tested. R1-substituents A–H are defined in Scheme 1b.
oxidatively cyclised with Pd(OAc)2/Cu(OAc)2 to render ketone 12
(Scheme 3). The oxime derivative of 12 underwent a Beckmann15
rearrangement to deliver 13 (R1 = Cl). Suzuki couplings led to the
styrene derivatives 13B and 13C, the phenyl analogues 13F and
13I and the pyridyl substituted tetracycles 13J and 13K. 14F—a tet-
racycle with an annulated cyclopentanone—was obtained from 12
via Suzuki coupling.
and leads to a weak MK2 inhibition (IC50 = 0.9
2-fluorophenyl group of 10G stays nearly in plane with the tetracy-
cle and remains a potent MK2 inhibitor (IC50 = 0.085 M). Pyridyl
derivatives 10J and 10K were the most potent MK2 inhibitors
(IC50 = 0.005 M) of the above series with a still modest 15- and
sevenfold selectivity over JNK2. Consequently, their cellular effi-
cacy may not be exclusively MK2-driven. N-Methylation of the lac-
tam—exemplified by 11A—was not tolerated. This is in agreement
with the finding, that the lactam-NH binds to Asp207 (see X-ray
analysis, Fig. 2).
lM), while the small
l
l
The styrene derivatives 13B and 13C (Table 3) were potent MK2
inhibitors (IC50 60.005
lM), but unselective as expected. The phe-
nyl derivatives 13F and 13I were potent MK2 inhibitors (IC50
60.016
l
M) and displayed good selectivity. Compound 13F
Tetracycles 13 with a six-membered lactam-ring were prepared
from 3-chloro-5-aminoisoquinoline and cyclopentane-1,3-dione,
which were condensed to an enaminone intermediate and then
showed
a
100-fold selectivity over JNK2, potently inhibited
hsp27 phosphorylation (IC50 = 0.5 lM) and the LPS induced release
N
N
N
a)b)
O
O
Cl
Cl
N
N
H
NH2
3
c)d)
h)f)g)i)
e)f)g)
N
O
O
O
R1
Cl
R1
O
Br
OH
N
H
N
O
NH
O
N
H
O
HN
5 (R2=H)
4
7
R2
6 (R2=Me)
j)g)i)f)
N
O
R1
NH
N
N
H
8
Scheme 1a. Reagents and conditions: (a) 3-oxobutyric acid tert-butyl ester, HOAc, 50 °C, 4 h, 66%; (b) Pd(OAc)2, Cu(OAc)2, DMF, 120 °C, 10 min, 63%; (c) BOC2O, DMAP,
diethylene glycol dimethylether, 120 °C, 30 min, 97%; (d) NBS, dibenzoyl peroxide, CCl4, reflux 2.5 h, 91%; (e) dioxane, MeNH2 or NH3concd microwave, 100 °C, 15 min, 60–70%;
(f) R1–B(OH)2 or pinacol ester, PdCl2(dppf), K2CO3, DMF/H2O, 90 °C, 1.5 h, 40–45%; (g) HClconcd 2 min, 25 °C; (h) tert-butyl N-hydroxycarbamate, K2CO3, dioxane, reflux
15 min., 65%; (i) EDCI, HOBt, DMF, 25 °C, 4.5 h, 55%; (j) methylhydrazine, NaHCO3, dioxane, reflux 3 h.

L. Revesz et al. / Bioorg. Med. Chem. Lett. 20 (2010) 4715–4718
4717
a)b)
c)d)e)
N
N
N
O
O
R1
Cl
Cl
NH
NH2
N
H
N
H
13
12
e)
N
O
R1
N
H
14
Scheme 3. Reagents and conditions: (a) 3-chloro-5-aminoisoquinoline and cyclo-
pentane-1,3-dione heated as melt at 120 °C for 25 min; (b) Pd(OAc)2, Cu(OAc)2,
DMF, 120 °C, 25 min., 88%; (c) NH2OHꢁHCl, pyridine, reflux 3 h, 70%. d) PPA, 130 °C,
75 min, 70% (e) R1–B(OH)2 or pinacol ester PdCl2(PPh3)2, 2 N Na2CO3, 1-propanol,
microwave, 160 °C, 15 min., 25–56%. R1-substituents A–H are defined in Scheme 1b.
Table 3
Compd MK2a
M)
TNFb
M)
p-hsp27c
M)
JNK2d
M)
Selectivityd
(
l
(
l
(
l
(
l
13B
13C
13F
13I
13J
13K
14F
0.001
0.005
0.015
0.016
0.010
0.012
0.160
0.009
0.032
0.097
0.294
0.283
0.260
1.400
nt
nt
0.198
0.076
1.42
0.269
0.418
0.649
2.200
13
8
0
3
2
0.5
1.3
2.5
0.7
5.8
Figure 2. Crystal structure of 13K bound to MK2.16
2
0
a)b)c)
Cl
d)
N
N
N
O
O
IC50 values are reported as the mean of P2 experiments with a standard deviation
of less than 50%.
O
R1
Cl
O
N
H
N
H
N
a-d
N
N
see Table 1.
R2
O
R2
Br
O
10 (R2=H)
4
9
1,3-dione in analogy to the tetracycles with a six-membered lac-
tam-ring described above (Scheme 4).
11 (R2=Me)
Scheme 2. Reagents and conditions: (a) dioxane, MeNH2 or NH3concd microwave,
100 °C, 15 min, 60–70%; (b) HClconcd 2 min, 25 °C; (c) EDCI, HOBt, DMF, 25 °C; (d)
R1–B(OH)2 or pinacol ester, PdCl2(dppf), K2CO3, DMF/H2O, 90 °C, 1.5 h, 40–45%. For
10D: 3-fluoroaniline, PdCl2(dppf), R(+)-BINAP, 2-dicyclohexyl-phosphino-20-(N,N-
dimethylamino)-biphenyl, Cs2CO3, DMF, microwave, 160 °C, 4 h, 34%. R1-substitu-
ents A–H are defined in Scheme 1b.
Styrene substituted tetracycles 16A–C (Table 4) were weak
MK2 inhibitors (IC50 60.253 lM) and unselective as expected. Phe-
nyl substituted tetracycles 16E and 16I were not much weaker
MK2 inhibitors than their five- and six-membered lactam ana-
logues, but displayed weak cellular activity. Pyridyl analogues
16J and 16K were ꢀ8-fold weaker against MK2 than six-membered
lactam analogues 13J, 13K and showed almost no cellular activity.
The lactam-ring size of the tetracycles determines their relative
potency at MK2, with the six-membered lactams being more
potent than the five-membered lactams, the latter being more po-
tent than the seven-membered lactams. R1-substituents had a
of TNF
less potent against MK2 (IC50 = 0.16
Pyridyl substituted tetracycles 13J and 13K were potent MK2
inhibitors (IC50 60.012 M). 13K showed a 54-fold preference for
MK2 over JNK2, inhibited hsp27 phosphorylation (IC50 = 0.7 M)
and LPS induced TNF release (IC50 = 0.26 M). The cellular effica-
a
(IC50 = 0.197
l
M) from hPBMCs. Ketone 14F was ꢀ10-fold
l
M) and similarly selective.
l
l
a
l
N
N
N
a)b)
c)d)e)
O
O
cies of both 13F and 13K are therefore linked with high probability
to MK2 inhibition.
R1
Cl
Cl
NH
N
H
NH2
N
H
Tetracycles 16 with
a
seven-membered lactam-ring were
16
15
prepared from 3-chloro-5-aminoisoquinoline and cyclohexane-
Scheme 4. Reagents and conditions: (a) 3-chloro-5-aminoisoquinoline and cyclo-
hexane-1,3-dione heated as melt at 120 °C for 25 min; (b) Pd(OAc)2, Cu(OAc)2, DMF,
120 °C, 25 min, 37%; (c) NH2OHꢁHCl, pyridine, reflux 3 h, 70%; (d) PPA, 130 °C,
75 min., 70%; (e) R1–B(OH)2 or pinacol ester PdCl2(PPh3)2, 2 N Na2CO3, 1-propanol,
microwave, 160 °C, 15 min., 29–63%. R1-substituents A–H are defined in Scheme 1b.
Table 2
Compd MK2a
M)
TNFb
M)
p-hsp27c
M)
JNK2d
M)
Selectivityd
(
l
(
l
(
l
(
l
Table 4
10A
10B
10C
10D
10E
10G
10H
10I
0.010
0.008
0.010
0.073
0.027
0.085
0.900
0.032
0.005
0.005
1.189
0.014
0.005
0.014
0.015
0.198
0.912
nt
0.280
0.556
0.416
nt
nt
nt
nt
nt
nt
5.078
nt
1.28
9.33
1.68
nt
0.002
0.022
0.034
0.001
nt
0.030
nt
0.011
0.075
0.038
nt
9
Compd MK2a
M)
TNFb
M)
p-hsp27c
M)
JNK2d
M)
Selectivityd
19
15
12
nt
nt
nt
6
1
3
nt
(
l
(
l
(
l
(
l
16A
16B
16C
16E
16I
0.116
0.253
0.172
0.072
0.167
0.080
0.096
0.202
0.140
>1
0.580
>3
nt
nt
nt
nt
nt
0.085
0.980
0.700
nt
nt
3.170
2.700
8
4
5
1
0
0
0
10J
10K
11A
16J
16K
8.6
8.38
>30
8.3
IC50 values are reported as the mean of P2 experiments with a standard deviation
IC50 values are reported as the mean of P2 experiments with a standard deviation
of less than 50%.
of less than 50%.
a-d
See Table 1.
a-d
See Table 1.

4718
L. Revesz et al. / Bioorg. Med. Chem. Lett. 20 (2010) 4715–4718
5. (a) Anderson, D. R.; Hegde, S.; Reinhard, E.; Gomez, L.; Vernier, W. F.; Lee, L.;
Liu, S.; Sambandam, A.; Snider, P. A.; Masih, L. Bioorg. Med. Chem. Lett. 2005, 15,
1587; (b) Wu, J.-P.; Wang, J.; Abeywardane, A.; Andersen, D.; Emmanuel, M.;
Gautschi, E.; Goldberg, D. R.; Kashem, M. A.; Lukas, S.; Mao, W.; Martin, L.;
Morwick, T.; Moss, N.; Pargellis, C.; Patel, U. R.; Patnaude, L.; Peet, G. W.; Skow,
D.; Snow, R. J.; Ward, Y.; Werneburg, B.; White, A. Bioorg. Med. Chem. Lett. 2007,
17, 4664; (c) Trujillo, J. I.; Meyers, M. J.; Anderson, D. R.; Hegde, S.; Mahoney, M.
W.; Vernier, W. F.; Buchler, I. P.; Wu, K. K.; Yang, S.; Hartmann, S. J.; Reitz, D. B.
Bioorg. Med. Chem. Lett. 2007, 17, 4657; (d) Goldberg, D. R.; Choi, Y.; Cogan, D.;
Corson, M.; DeLeon, R.; Gao, A.; Gruenbaum, L.; Hao, M. H.; Joseph, D.; Kashem,
M. A.; Miller, C.; Moss, N.; Netherton, M. R.; Pargellis, C. P.; Pelletier, J.; Sellati,
R.; Skow, D.; Torcellini, C.; Tseng, Y.-C.; Wang, J.; Wasti, R.; Werneburg, B.; Wu,
J. P.; Xiong, Z. Bioorg. Med. Chem. Lett. 2008, 18, 938; (e) Xiong, Z.; Gao, D. A.;
Cogan, D. A.; Goldberg, D. R.; Hao, M.-H.; Moss, N.; Pack, E.; Pargellis, C.; Skow,
D.; Trieselmann, T.; Werneburg, B.; White, A. Bioorg. Med. Chem. Lett. 2008, 18,
1994; (f) Anderson, D. R.; Meyers, M. J.; Vernier, W. F.; Mahoney, M. W.;
Kurumbail, R. G.; Caspers, N.; Poda, G. I.; Schindler, J. F.; Reitz, D. B.; Mourey, R.
J. J. Med. Chem. 2007, 50, 2647; (g) Lin, S.; Lombardo, M.; Malkani, S.; Hale, J. J.;
Mills, S. G.; Chapman, K.; Thompson, J. E.; Zhang, W. X.; Wang, R.; Cubbon, R.
M.; O’Neill, E. A.; Luell, S.; Carballo-Jane, E.; Yang, L. Bioorg. Med. Chem. Lett.
2009, 19, 3238; (h) Anderson, D. R.; Meyers, M. J.; Kurumbail, R. G.; Caspers, N.;
Poda, G. I.; Long, S. A.; Pierce, B. S.; Mahoney, M. W.; Mourey, R. J. Bioorg. Med.
Chem. Lett. 2009, 19, 4878; (i) Anderson, D. R.; Meyers, M. J.; Kurumbail, R. G.;
Caspers, N.; Poda, G. I.; Long, S. A.; Pierce, B. S.; Mahoney, M. W.; Mourey, R. J.;
Parikh, M. D. Bioorg. Med. Chem. Lett. 2009, 19, 4882; (j) Harris, C. M.; Ericsson,
A. M.; Argiriadi, M. A.; Barberis, C.; Borhani, D. W.; Burchat, A.; Calderwood, D.
J.; Cunha, G. A.; Dixon, R. W.; Frank, K. E.; Johnson, E. F.; Kamens, J.; Kwak, S.; Li,
B.; Mullen, K. D.; Perron, D. C.; Wang, L.; Wishart, N.; Wu, X.; Zhang, X.; Zmetra,
T. R.; Talanian, R. V. Bioorg. Med. Chem. Lett. 2010, 20, 334.
modulating effect on potency and selectivity. High selectivity was
conferred by phenyl and pyridyl substituents, while styryl substit-
uents and anilines showed affinity to off-target kinases such as
JNK2. Insight into the binding mode of tetracycles was obtained
by X-ray crystallography of 13K with MK2 (47–364). 13K bound
to the hinge region with a classical H-bond formed between the
N-atom of its pyridine and the backbone amide nitrogen of
Leu141. The pyrrole NH of 13K bound to a water molecule, which
was H-bonding to the backbone carbonyl of Leu70. The lactam car-
bonyl showed H-bonding to Lys93, the lactam N–H bound to
Asp207 (Fig. 2).
Several tetracyclic lactams were tested17 in vivo, but none
showed oral efficacy. The tetracyclic ketone 14F however proved
to be orally bioavailable and inhibited 73% of LPS induced TNF
lease in mice with plasma levels reaching 13
M after 2 h.17 Since
14F was shown to inhibit LPS induced release of TNF from
hPBMCs (Table 3) and hsp27 phosphorylation with IC50 values of
1.4 M and 5.8 respectively, one may assume, that this
a re-
l
a
l
lM
in vivo effect is MK2-driven.
N
F
O
N
H
O
6. Schlapbach, A.; Feifel, R.; Hawtin, S.; Heng, R.; Koch, G.; Moebitz, H.; Revesz, L.;
Scheufler, C.; Velcicky, J.; Waelchli, R.; Huppertz, C. Bioorg. Med. Chem. Lett.
2008, 18, 6142.
14F
7. Velcicky, J.; Feifel, R.; Hawtin, S.; Heng, R.; Huppertz, C.; Koch, G.; Kroemer, M.;
Moebitz, H.; Revesz, L.; Scheufler, C.; Schlapbach, A. Bioorg. Med. Chem. Lett.
2010, 20, 1293.
8. Schlapbach, A.; Huppertz, C. Future Med. Chem. 2009, 1, 1243–1257.
9. Iida, H.; Yuasa, Y.; Kibayashi, C. Journal. Org. Chem. 1980, 45, 2938.
10. Suzuki, A. Pure Appl. Chem. 1991, 63, 419.
Further optimization of these compound classes for oral bioavail-
ability is ongoing.
Acknowledgements
11. Revesz, L.; Schlapbach, A.; Waelchli, R. WO 2008025512. Chem. Abstr. 2008,
148, 331658.
12. In-house off-target screen for MK2 inhibitors: ALK, CDK2A, ERK2, EphA4, FGFR-
4, FGFR3K, HER1, HER2, INS1R, JAK1, JAK2, KDR, LCK, PDGFRa, PDK1, PKA, PLK1,
RET, SYK, cABL, cABLT315, cKIT, cMET, p38a, JNK1, JNK2.
13. Gaillard, P.; Jeanclaude-Etter, I.; Ardissone, V.; Arkinstall, S.; Cambet, Y.;
Camps, M.; Chabert, C.; Church, D.; Cirillo, R.; Gretener, D.; Halazy, S.; Nichols,
A.; Szyndralewiez, C.; Vitte, P.-A.; Gotteland, J.-P. Journal. Med. Chem. 2005, 48,
4596.
The kilolab and its coworkers are gratefully acknowledged for
preparing ample quantities of 3-chloro-5-aminoisoquinoline. We
thank Dr. Karl Welzenbach for technical assistance in measuring
phosphorylation of hsp27 using flow cytometry.
14. Hartwig, J. F. Acc. Chem. Res. 2008, 41, 1534.
15. Gawley, R. E. Org. React. 1988, 35, 1.
References and notes
16. The X-ray coordinates are deposited with RCSB Protein Data Bank, deposition
code is 3M42. The protein used in this study is a segment of MK2 containing
residues 47–364d(216–237)G.
17. MK2 inhibitors (100 mg/kg po) were administered to OFI mice (female,
8 weeks old), followed by LPS injection (20 mg/kg) 1 h later. One hour post
LPS injection the experiment was terminated and blood withdrawn.
Compound blood levels were determined by LC–MS/MS and plasma levels of
1. Chen, Z.; Gibson, T. B.; Robinson, F.; Silvestro, L.; Pearson, G.; Xu, B.; Wright, A.;
Vanderbilt, C.; Cobb, M. H. Chem. Rev. 2001, 101, 2449.
2. Gaestel, M. Nat. Rev. Mol. Cell. Biol. 2006, 7, 120.
3. Kotlyarov, A.; Yannoni, Y.; Fritz, S.; Laass, K.; Telliez, J. B.; Pitman, D.; Lin, L. L.;
Gaestel, M. Mol. Cell Biol. 2002, 22, 4827.
4. Hegen, M.; Gaestel, M.; Nickerson-Nutter, C. L.; Lin, L.-L.; Telliez, J.-B. J.
Immunol. 2006, 177, 1913.
mouse TNFa determined by ELISA.