M. L. Curtin et al. / Bioorg. Med. Chem. Lett. 22 (2012) 4750–4755
4755
5. For a review of the most recently approved KDR inhibitor, see: Commander, H.;
Whiteside, G.; Perry, C. Drugs 2011, 71, 1355.
6. Fojo, T. Oncologist 2008, 13, 277.
2500
2000
1500
1000
500
Vehicle
Cmpd 1 (50 mg/kg, po)
Cmpd 17 (10 mg/kg, ip)
Cmpd (+)-18 (10 mg/kg, ip)
Cmpd (+)-32 (10 mg/kg, ip)
Cmpd 37 (10 mg/kg, ip)
Cmpd 39 (10 mg/kg, ip)
7. Dai, Y.; Hartandi, K.; Ji, Z.; Ahmed, A. A.; Albert, D. H.; Bauch, J. L.; Bouska, J. J.;
Bousquet, P. F.; Cunha, G. A.; Glaser, K. B.; Harris, C. M.; Hickman, D.; Guo, J.; Li,
J.; Marcotte, P. A.; Marsh, K. C.; Moskey, M. D.; Martin, R. L.; Olson, A. M.;
Osterling, D. J.; Pease, L. P.; Soni, N. B.; Stewart, K. D.; Stoll, V. S.; Tapang, P.;
Reuter, D. R.; Davidsen, S. K.; Michaelides, M. R. J. Med. Chem. 2007, 50, 1584.
8. Curtin, M. L.; Heyman, H. R.; Frey, R. R.; Soni, N. B.; Marcotte, P. A.; Pease, L. J.;
Glaser, K. B.; Magoc, T. J.; Tapang, P.; Albert, D. H.; Osterling, D. J.; Olson, A. M.;
Bouska, J. J.; Guan, Z.; Martin, R. L.; Preusser, L. C.; Polakowsi, J. S.; Stewart, K.
D.; Tse, C.; Davidsen, S. K.; Michaelides, M. R. Bioorg. Med. Chem. Lett. 2012, 22,
3208.
9. (a) McLaughlin, J.; Markovtsov, V.; Li, H.; Wong, S.; Gelman, M.; Zhu, Y.; Franci,
C.; Lang, D. W.; Pali, E.; Lasaga, J.; Low, C.; Zhao, F.; Chang, B.; Gururaja, T. L.; Xu,
W.; Baluom, M.; Sweeney, D.; Carroll, D.; Sran, A.; Thota, S.; Parmer, M.;
Romane, A.; Clemens, G.; Grossbard, E.; Qu, K.; Jenkins, Y.; Kinoshita, T.; Taylor,
V.; Holland, S. J.; Argade, A.; Singh, R.; Pine, P.; Payan, D. G.; Hitoshi, Y. J. Cancer
Res. Clin. Oncol. 2010, 136, 99; (b) To our knowledge, SAR data for AS703569 has
not been disclosed.
0
/ /
20
25
30
35
Days
Dosing schedule
10. KDR and Aurora
B enzymatic IC50 values were determined using a
Figure 3. DoHH-2 mouse tumor growth model. All compounds dosed q7dx3
(n = 10, SCID).
homogeneous time-resolved fluorescence (HTRF) kinase assay with an ATP
concentration of 1 mM as described in Ref. 11.
11. Dai, Y.; Guo, Y.; Frey, R. R.; Ji, Z.; Curtin, M. L.; Ahmed, A. A.; Albert, D. H.;
Arnold, L.; Barlozzari, T.; Bauch, J. L.; Bouska, J. J.; Bousquet, P. F.; Cunha, G. A.;
Glaser, K. B.; Guo, J.; Li, J.; Marcotte, P. A.; Marsh, K. C.; Moskey, M. D.; Pease, L.
J.; Stewart, K. D.; Stoll, V. S.; Tapang, P.; Wishart, N.; Davidsen, S. K.;
Michaelides, M. R. J. Med. Chem. 2005, 48, 6066.
12. Aurora B cellular activity was assessed by measuring induction of polyploidy in
the NCI-H1229 (NSCLC) cell line as described in Ref. 13 Initial dose–response
data obtained with this assay indicated that maximal polyploid response to
inhibitors typically did not exceed 50% and was associated with apoptosis and
loss in cell number. To avoid this complexity, a threshold potency, defined as
the concentration necessary to produce DNA content >4 N in 15% of the cell
Table 5
Partial kinase inhibition profile of 17
Kinase
IC50 (nM)a
Kinase
IC50 (nM)a
KDR
AurA
Flt3
GSK3b
PDGFR
6
3
1
34
1
Lck
FGFR2
Syk
PKC
Wee1
7
2
44
82
2800
population (EC15), was chosen rather than the more common EC50
.
e
13. McClellan, W. J.; Dai, Y.; Abad-Zapatero, C.; Albert, D. H.; Bouska, J. J.; Glaser, K.
B.; Magoc, T. J.; Marcotte, P. A.; Osterling, D. J.; Stewart, K. D.; Davidsen, S. K.;
Michaelides, M. R. Bioorg. Med. Chem. Lett. 2011, 21, 5620.
14. The KDR cellular IC50 values represent inhibition of KDR phosphorylation in
NIH3T3 cells stably transfected with full length human KDR. The procedure
was followed as described in Ref. 11.
a
a
TR-FRET assay.
15. Compounds were dissolved in DMSO (10 mM), added to pH 7.2 buffer (1:100
dilution) and equilibrated for 24 h with vigorous shaking. The samples were
centrifuged and compound concentration in the supernatant was determined
using chemiluminescent nitrogen detection (CLND).
16. After this work was completed, the FDA approved the first dianilinopyrimidine
RTK inhibitor, pazopanib, for the treatment of renal cell carcinoma. For an
account of the discovery of this compound, see: Harris, P. A.; Boloor, A.;
Cheung, M.; Kumar, R.; Crosby, R. M.; Davis-Ward, R. G.; Epperly, A. H.; Hinkle,
K. W.; Hunter, R. N.; Johnson, J. H.; Knick, V. B.; Laudeman, C. P.; Luttrell, D. K.;
Mook, R. A.; Nolte, R. T.; Rudolph, S. K.; Szewczyk, J. R.; Truesdale, A. T.; Veal, J.
M.; Wang, L.; Stafford, J. A. J. Med. Chem. 2008, 51, 4632.
17. All intermediates and final compounds are racemates unless otherwise noted.
18. (1S,2S,3R,4R)-Carboxamide 7 was prepared using enzymatic chiral resolution
of lactam 6 as described in WO2006/055561.
19. A homology model of human Aurora B kinase was constructed using template
PDB code 1ATP for protein kinase A. The ligand orientation of (+)-38 was
guided by using template 2NP8 for Aurora A Kinase according to the procedure
described in Ref. 11.
aqueous solubility and moderate antitumor activity in a mouse tu-
mor model after weekly ip dosing. Unfortunately these compounds
were pan-kinase inhibitors that suffered from narrow therapeutic
indices which prohibited their use as antitumor agents.
References and notes
1. Marumoto, T.; Zhang, D.; Saya, H. Nat. Rev. Cancer 2005, 5, 42.
2. Matthews, N.; Visintin, C.; Hartzoulakis, B.; Jarvis, A.; Selwood, D. L. Expert Rev.
Anticancer Ther. 2006, 6, 109.
3. (a) Yang, H.; Burke, T.; Dempsey, J.; Diaz, B.; Collins, E.; Toth, J.; Beckmann, R.;
Ye, X. FEBS Lett. 2005, 579, 3385; (b) Girdler, F.; Gascoigne, K. E.; Eyers, P. A.;
Hartmuth, S.; Crafter, C.; Foote, K. M.; Keen, N. J.; Taylor, S. S. J. Cell Sci. 2006,
119, 3664.
4. Ferrara, N.; Gerber, H. P.; LeCouter, J. Nat. Med. 2003, 9, 669.