6994
S. A. Mitchell et al. / Bioorg. Med. Chem. Lett. 19 (2009) 6991–6995
Table 7
Linker functionality survey
sessed in Phase I clinical trials in oncology. It is envisioned that
potent simultaneous inhibition of these RTKs may prove to be a
successful combination for the treatment of certain cancer types.
References and notes
1. (a) Surawska, H.; Ma, P. C.; Salgia, R. Cytokine Growth Factor Res. 2004, 15, 419;
(b) Brantley-Sieders, D. M.; Chen, J. Angiogenesis 2004, 7, 17; (c) Cheng, N.;
Brantley, D. M.; Chen, J. Cytokine Growth Factor Res. 2002, 13, 75; (d) Dodelet, V.
C.; Pasquale, E. B. Oncogene 2000, 19, 5614; (e) Sullivan, D. C.; Bicknell, R. Br. J.
Cancer 2003, 89, 228.
2. (a) Davalos, V.; Dopeso, H.; Castano, J.; Wilson, A. J.; Vilardell, F.; Romero-
Gimenez, J.; Espin, E.; Armengol, M.; Capella, G.; Mariadason, J. M.; Aaltonen, L.
A., ; Schwartz, S., Jr. Cancer Res. 2006, 66, 8943; (b) Kumar, S. R.; Singh, J.; Xia,
G.; Krasnoperov, V.; Hassanieh, L.; Ley, E. J.; Scehnet, J.; Kumar, N. G.; Hawes,
D.; Press, M. F.; Weaver, F. A.; Gill, P. S. Am. J. Pathol. 2006, 169, 279.
3. Noren, N. K.; Lu, M.; Freeman, A. L.; Koolpe, M.; Pasquale, E. B. Proc. Natl. Acad.
Sci. 2004, 101, 5583.
4. (a) Folkman, J. N. Eng. J. Med. 1971, 285, 1182; (b) Ferrara, N.; Kerbel, R. S. Nature
2005, 438, 967; (c) Clarke, S. J.; Sharma, R. Aust. Prescr. 2006, 29, 9; (d)
Mazitschek, R.; Giannis, A. Curr. Opin. Chem. Biol. 2004, 8, 432.
5. Fernando, N. H.; Hurwitz, H. I. Oncologist 2004, 9, 11; Hurwitz, H. I.;
Fehrenbacher, L.; Hainsworth, J. D.; Heim, W.; Berlin, J.; Holmgren, E.;
Hambleton, J.; Novotny, W. F.; Kabbinavar, F. J. Clin. Oncol. 2005, 23, 3502.
6. Wilhelm, S.; Carter, C.; Lynch, M.; Lowinger, T.; Dumas, J.; Smith, R. A.;
Schwartz, B.; Simantov, R.; Kelley, S. Nat. Rev. Drug Discovery 2006, 5, 835.
7. Mendel, D. B.; Laird, A. D.; Xin, X.; Louie, S. G.; Christensen, J. G.; Li, G.; Schreck,
R. E.; Abrams, T. J.; Ngai, T. J.; Lee, L. B.; Murray, L. B.; Carver, J.; Chan, E.; Moss,
K. G.; Haznedar, J. Ö.; Sukbuntherng, J.; Blake, R. A.; Sun, L.; Tang, C.; Miller, T.;
Shirazian, S.; McMahon, G.; Cherrington, J. M. Clin. Cancer Res. 2003, 9, 327.
8. (a) Martinelli, E.; Troiani, J.; Morgillo, F.; Piccirillo, M. C.; Monaco, K.; Morelli,
M. P.; Cascone, T.; Ciardiello, F. Target. Oncol. 2006, 1, 123; (b) Smith, J. K.;
Mamoon, N. M.; Duhé, R. J. Oncol. Res. 2004, 14, 175; (c) Wedge, S. R.; Kendrew,
J.; Hennequin, L. F.; Valentine, P. J.; Barry, S. T.; Brave, S. R.; Smith, N. R.; James,
N. H.; Dukes, M.; Curwen, J. O.; Chester, R.; Jackson, J. A.; Boffey, S. J.; Kilburn, L.
L.; Barnett, S.; Richmond, G. H. P.; Wadsworth, P. F.; Walker, M.; Bigley, A. L.;
Taylor, S. T.; Cooper, L.; Beck, S.; Jürgensmeier, J. M.; Ogilvie, D. J. Cancer Res.
2005, 65, 4389; (d) Polverino, A.; Coxon, A.; Starnes, C.; Diaz, Z.; DeMelfi, T.;
Wang, L.; Bready, J.; Estrada, J.; Cattley, R.; Kaufman, S.; Chen, D.; Gan, Y.;
Kumar, G.; Meyer, J.; Neervannan, S.; Alva, G.; Talvenheimo, J.; Montestruque,
S.; Tasker, A.; Patel, V.; Radinsky, R.; Kendall, R. Cancer Res. 2006, 66, 8715.
9. Wakelee, H. A.; Adjei, A. A.; Halsey, J.; Dugay, J. D.; Hanson, L. J.; Reid, J. M.;
Piens, J. R.; Sikic, B. I. Poster 3044, American Society of Clinical Oncology,
Atlanta, GA, 2006, June 2–6.
Compds
L
EphB4 IC50 (lM)
2
70
71
72
73
74
75
76
77
3-NHCONH-
3-NHCO–
3-NHCOCH2–
3-NHCONHCH2–
3-NHSO2–
3-CH2NHCONH–
3-CONHCH2–
3-NHCH2CONH–
4-NHCONH–
0.062
0.744
2.4
2.1
50
6.8
11.1
2.9a
50
a
2-OMe–5-CF3-phenyl urea instead of 3-CF3-phenyl.
10. (a) Marx, J. Science 2005, 308, 1248; (b) Kerbel, R. S. Science 2006, 312, 1171; (c)
Rosen, L. S. Oncolog. J. 2006, 10, 382; (d) Ranieri, G.; Patruno, R.; Ruggieri, E.;
Montemurro, S.; Valero, P.; Ribatti, D. Curr. Med. Chem. 2006, 13, 1845; (e)
Shinkaruk, S.; Bayle, M.; Lain, G.; Deleris, G. Curr. Med. Chem. Anticancer Agents
2003, 3, 95; (f) Underiner, T. L.; Ruggieri, B.; Gingrich, D. E. Curr. Med. Chem.
2004, 11, 731.
Figure 3. Docking of 2 in EphB4 homology model.
11. (a) Schwede, W.; Jaroch, S.; Bader, B.; Hillig, R.; Ter Laak, A.; Zopf, D. PCT Int.
Appl. WO 06/066955, 2006.; (b) Schwede, W.; Kuenzer, H.; Bader, B.; Hillig, R.;
Ter Laak, A.; Moenning, U.; Schmidt, A.; Zopf, D. PCT Int. Appl. WO 06/063805,
2006.; (c) Herz, T.; Krauss, R.; Kubbutat, M.; Lang, M.; Schaechtele, C.; Tasler, S.;
Totzke, F. U.S. Patent Appl. US 06/135782, 2006.; (d) Schmiedeberg, N.; Furet,
P.; Imbach, P.; Holzer, P. PCT Int. Appl. WO 06/050946, 2006.; (e) Berset, C.;
Audetat, S.; Tietz, J.; Gunde, T.; Barberis, A.; Schumacher, A.; Traxler, P. PCT Int.
Appl. WO 05/120513, 2005.; (f) Yoshiaki, W. PCT Int. Appl. WO 05/049576,
2005.; (g) Armistead, D. M.; Bemis, J. E.; Buchanan, J. L.; Dipietro, L. V.; Elbaum,
D.; Geuns-Meyer, S. D.; Habgood, G. J.; Kim, J. L.; Marshall, T. L.; Novak, P. M.;
Nunes, J. J.; Patel, V. F.; Toledo-Sherman, L. M.; Zhu, X. U.S. Patent Appl. US 04/
116388, 2004.; (h) Miyazaki, Y.; Nakano, M.; Sato, H.; Truesdale, A. T.; Stuart, J.
D.; Nartey, E. N.; Hightower, K. E.; Kane-Carson, L. Bioorg. Med. Chem. Lett. 2006.
Table 8
Biochemical multiplex activity of lead compounds IC50 (nM)
Compds
EphB4
VEGFR2
Tie2
45
46
59
37
49
26
7
13
10
33
37
75
12. Miyazaki, Y.; Nakano, M.; Sato, H.; Truesdale, A. T.; Stuart, J. D.; Nartey, E. N.;
Hightower, K. E.; Kane-Carson, L. Bioorg. Med. Chem. Lett. 2007, 17, 250.
13. (a) Wilks, A. F.; Bu, X.; Burns, C.J. PCT Int. Appl. WO 03/099811, 2003.; (b) Guzi,
T. J.; Paruch, K.; Dwyer, M. P.; Zhao, L.; Curran, P. J.; Belanger, D. B.; Hamann, B.;
Reddy, P. A. Siddiqui, M. A. U.S. Patent Appl. US 06/106023, 2006.
onstrated potent inhibition of EphB4, VEGFR2 and Tie2 autophos-
phorylation, as shown in Table 9.
These analogs represent one of the first classes of small mole-
cule kinase inhibitors that display the triplet activity profile against
EphB4, VEGFR2 and Tie2. We have identified a subsequent class of
molecules that potently inhibits a triplet set of angiogenesis-re-
lated kinases, and a member from that group is currently being as-
14. EphB4 biochemical assay: Dilutions of compounds are made in 100% DMSO at
20X the final desired concentration. Compounds in 100% DMSO are transferred
(1.25
containing the final concentrations (in 25
kinase buffer, 0.5 M PTK biotinylated peptide substrate 2, and 18.6 ng/well
ng/well of EphB4 kinase is added to all wells, except the four negative control
wells (which contain no kinase), and mixed. To initiate the reaction, 5 L of
550 M ATP is added to each well. (final concentration of ATP = 110 M). The
reactions are incubated for 1 h at room temperature (rt). After incubation a
quantity of 8.35 L of a 4X SA-APC detection mix is added to each well. The
l
L) to the 96 well assay plate.
A
18.75
lL volume of master mix
lL) of 0.01% BSA, 1X cell signaling
l
l
l
l
Table 9
Cellular multiplex activity of lead compounds IC50 (nM)
l
final concentration of Eu-labelled PT66 antibody is 1 nM and the SA-APC is
20 nM (based on the SA moiety). The reaction plates are incubated at rt for at
least 15 min after SA-APC detection mix addition. The reaction plates are read
on an Envision plate reader (Perkin–Elmer) with 605 nm excitation at 605 nm
and 640 nm emission wavelengths. Values are corrected for the fluorescence in
Compds
EphB4
VEGFR2
Tie2
45
46
59
50
400
50
100
100
100
50
100
250