J. D. Hansen et al. / Bioorg. Med. Chem. Lett. 18 (2008) 4692–4695
4695
Table 4
Table 5
B-Raf activity of C4-modified pyrazoles
In vitro stability, permeability, and solubility
R
2
10
7
18
31
N
CH3
Hepat. Cla
Caco-2
7
6
8.5
High
3c
7.8
Med
52
8
High
590
N
High
High
N
Sol. @pH6.5b
4c
76c
a
Compound
R
B-Raf IC50 (nM)
pERK IC50 (nM)
20,000
a
Human hepatocyte clearance (mL/min/kg).
b
c
Aqueous thermodynamic solubility (
lg/mL).
Crystalline solid.
NH2
OH
30
18.4
N
NH
5000
ng/ml
180
160
140
120
100
80
31
32
33
34
35
36
<2.0
16.0
68.4
4.4
56
4500
N
OH
% control
4000
3500
3000
2500
2000
1500
1000
500
NH
10,000
—
N
OH
60
N
N
NH
40
18,000
1900
—
20
NH
0
0
1.5
NH
N
Figure 4. PK/PD evaluation of substituted pyrazoles after 25 mg/kg po dose in LOX
tumor-bearing mice. Four per group, error bars are SEM.
NH
68.4
70.0
O
References and notes
H
N
37
—
N
1. Hoshino, R.; Chantani, Y.; Yamori, T.; Tsuruo, T.; Oka, H.; Yoshida, O.; Shimada,
Y.; Ari-I, S.; Wada, H.; Fujimoto, J.; Kohno, M. Oncogene 1999, 18, 813.
2. (a) Davies, H.; Bignell, G. R.; Cox, C.; Stephens, P.; Edkins, S.; Clegg, S.; Teague, J.;
Woffendin, H.; Garnett, M. J.; Bottomley, W.; Davis, N.; Dicks, E.; Ewing, R.;
Floyd, Y.; Gray, K.; Hall, S.; Hawes, R.; Hughes, J.; Kosmidou, V.; Menzles, A.;
Mould, C.; Parker, A.; Stevens, C.; Watt, S.; Hooper, S.; Wilson, R.; Jayatilake, H.;
Gusterson, B. A.; Cooper, C.; Shipley, J.; Hargrave, D.; Pritchard-Jones, K.;
Maitland, N.; Chenevix-Trench, G.; Riggins, G. J.; Bigner, D. D.; Palmieri, G.;
Cossu, A.; Flanagan, A.; Nicholson, A.; Ho, J. W. C.; Leung, S. Y.; Yuen, S. T.;
Weber, B. L.; Seigler, H. F.; Darrow, T. L.; Paterson, H.; Marais, R.; Marshall, C. J.;
Wooster, R.; Stratton, M. R.; Futreal, P. A. Nature 2002, 417, 949; (b) Cohen, Y.;
Xing, M.; Mambo, E.; Guo, Z.; Wu, G.; Trink, B.; Beller, U.; Westra, W. H.;
Ladenson, P. W.; Sidransky, D. J. Natl. Cancer Inst. 2003, 95, 625; (c) Xu, X.;
Quiros, R. M.; Gattuso, P.; Ain, K. B.; Prinz, R. A. Cancer Res. 2003, 63, 4561.
3. Forbes, S.; Clements, J.; Dawson; Bamford, S.; Webb, T.; Dogan, A.; Flanagan,
A.; Teague, J.; Wooster; Futreal, P. A.; Stratton, M. R. Br. J. Cancer 2006, , 94,
318.
4. Wan, P. T.; Garnet, M. J.; Roe, S. M.; Lee, S.; Niculescu-Duvaz, D.; Good, V. M.;
Jones, C. M.; Marshall, C. J.; Springer, C. J.; Barford, D.; Marais, R. Cell 2004, 116,
855.
5. (a) Khazak, V.; Astsaturov, I.; Serebriiski, I.; Golemis, E. Expert Opin. Ther. Targets
2007, 11, 1587; (b) Li, N.; Batt, D.; Warmuth, M. Curr. Opin. Invest. Drugs 2007, 8,
452.
6. Tackle, A. K.; Brown, M. J. B.; Davies, S.; Dean, D. K.; Francis, G.; Gaiba, A.; Hird,
A. W.; King, F. D.; Lovell, P. J.; Naylor, A.; Reith, A. D.; Steadman, J. G.; Wilson, D.
M. Bioorg. Med. Chem. Lett. 2006, 16, 378 .
a
IC50 values reflect the average from at least three separate experiments.
Br
Br
a,b,c
NH
OH
O
d,e
N
N
N
N
N
38
39
40
N
HO
N
O
f,g
+ 40
O
B
N
N
OH
O
41
10
N
(GDC-0879)
(AR00341677)
7. ClinicalTrials.gov.; Identifier: NCT00304525.
Scheme 1. Reagents and conditions: (a) DMF–DMA, tol,
EtOH,
potassium carbonate, DMF; (e) water, MeOH, potassium carbonate,
tetrakis(triphenylphosphine)palladium, potassium carbonate, ACN–water,
D
, (78%); (b) hydrazine,
8. For assay description see: Laird, E.; Lyssikatos, J.; Welch, M.; Grina, J.; Hansen,
J.; Newhouse, B.; Olivero, A.; Topolav, G. WO 2006/084015 A2, 2006.
9. Coordinates for the B-Raf crystal structure have been deposited in PDB: 3d4q.
10. Compound 2 has an aqueous stability half life of 1.2 h at pH 1.2.
11. Bamborough, P.; Angell, R. M.; Bhamra, I.; Brown, D.; Bull, J.; Christopher, J. A.;
Cooper, A. W. J.; Fazal, L. H.; Giordano, I.; Hind, L.; Patel, V. K.; Ranshaw, L. E.;
Sims, M. J.; Skone, P. A.; Smith, K. J.; Vickerstaff, E.; Washington, M. Bioorg. Med.
Chem. Lett. 2007, 17, 4363.
D
,
(96%); (c) bromine, NaOAc, AcOH, (70%); (d) 2-bromoethyl acetate,
, (76%); (f)
, (74%);
D
D
(g) hydroxylamine hydrochloride, EtOH,
D, (78%).
Acknowledgement
12. Compound 39 has been described: Desbordes, P.; Guigues, F. WO 94/29300,
1994.
13. Detailed description of these assays can be found in the supporting information
from: Wallace, E. M.; Lyssikatos, J.; Blake, J. F.; Seo, J.; Yang, H. W.; Yeh, T. C.;
Perrier, M.; Jarski, H.; Marsh, V.; Poch, G.; Livingston, M. G.; Otten, J.; Hingorani,
G.; Woessner, R.; Lee, P.; Winkler, J.; Koch, K. J. Med. Chem. 2006, 49, 441.
The authors thank Andrew Allen for NMR analysis and Susan
Rhodes, Jennifer Otten, and Michelle Livingston for Caco, P450,
and solubility determinations.