1690
F. Pierre et al. / Bioorg. Med. Chem. Lett. 21 (2011) 1687–1691
tions. Additional characterization and optimization of our chemical
series will be the subject of future reports.43
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Figure 3. Effect of 7e in phase II (11–40 min) of the formalin response in mice. (h)
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Table 3
Antiviral activity of compound 14k against HIV-1 clinical isolates in fresh human
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Virus (subtype)
Endpoint
IC50
(
lM)
TC50
(lM)
Therapeutic
index TC50/IC50
00KNH1209 (A)
94US3393IN (B)
98IN022 (C)
RT
p24
RT
p24
RT
p24
RT
p24
RT
p24
RT
p24
RT
p24
0.40
0.17
0.54
0.19
0.26
0.20
0.24
0.13
0.08
0.14
0.28
0.39
0.24
0.10
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>248
>605
>184
>527
>387
>491
>419
>757
>1246
>726
>355
257
00KENKU3006 (D)
93TH073 (E)
93BR019 (BF)
G3 (G)
>410
>1018
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31. All compounds were characterized by LCMS (P95% pure). Selected compounds
were characterized by NMR. CK2 inhibition was measured in a radiometric
of pain induced flinching. However, 7e significantly reduced pain
during the phase II (11–40 min) in a dose-dependent manner com-
pared to vehicle treated mice, suggesting an anti-inflammatory
based mechanism. A single dose of morphine (3 mg/kg, subcutane-
ously injected) served as a positive control, and reduced formalin-
induced flinching in both phases of the test.
Preliminary anti-viral screening identified promising activity
for 14k against CCR5-tropic HIV-1 clinical isolates in human
peripheral blood mononuclear cells (PBMC). This compound was
further evaluated against a broader assortment of clinical isolates
(Table 3). Overall, compound 14k inhibited virus production at
sub micromolar IC50 values and had no cytotoxicity at high test
assay using human recombinant CK2 (aabb-holoenzyme) at [ATP] = 15 lM,
using the substrate peptide RRRDDDSDDD. The IC50 values were derived from
eight concentrations of test inhibitors.
concentration of 100
tic index.
lM in PBMC, providing a favorable therapeu-
32. Molecules were manually placed in the binding pocket (PDB structure 1JWH of
In summary, we have discovered multiple series of novel potent
pyrimido[4,5-c]quinolones, that act mechanistically as ATP-com-
petitive inhibitors of protein kinase CK2. Among these new chem-
ical entities, several representatives inhibited the protein with IC50
values below 10 nM. Molecular modeling and SAR suggested that
C-3 alkyl-amino substituted inhibitors such as 14k interacted
slightly differently with the hinge region of the protein, an obser-
vation that might potentially affect their kinase selectivity profile.
We have shown that representative analogs 7e and 14k were
active in cells, providing valuable tools for investigating the phar-
macology of CK2. Compound 14k showed in vitro anti-viral activity
with IC50 values as low as 80 nM against HIV-1 viruses with an
excellent therapeutic index. Orally available 7e demonstrated dose
dependent reduction of pain in vivo. These results confirm the
therapeutic potential of CK2 inhibitors, and suggest that properly
optimized inhibitors may deliver drugs for multiple disease indica-
human CK2a used as a template) and subjected to in situ minimization using
the CHARMm forcefield as implemented in Discovery Studio 2.5.1 from
Accelrys Inc.
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A.; Ruzzene, M. Cell Death Differ. 2005, 12, 668.
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36. Battistutta, R. Cell. Mol. Life Sci. 2009, 66, 1868.
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39. The sodium salt of 7e was administered as a DI water solution. The free acid of
14k was administered as a solution in water containing 0.4–0.8% L-arginine.
After administration in animals, blood samples were centrifuged and the
plasma was analyzed by LC–MS/MS technique. The pharmacokinetic
parameters were derived by noncompartmental analysis. Abbreviations: iv,
intravenous dose; po, oral dose; Cls, clearance; Vss, volume of distribution at