P. Eastwood et al. / Bioorg. Med. Chem. Lett. 21 (2011) 4130–4133
4133
Table 2
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ADME Profile for compound for 16
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Parameter
Value
% Turnover in human hepatic microsomesa
+NAPDH:
55
55
18(10)
3
ꢀNADPH:
CACO2: Papp (ꢂ10ꢀ6 cm/s)b AꢀB (BꢀA):
11. Ferraccioli, G. F. Curr. Opin. Anti-Inflammatory Immunomodul. Invest. Drugs
2000, 2, 74.
Thermodymanic solubilityc
(lg/mL at pH 7.4):
12. Haddad, J. J. Curr. Opin. Investig. Drugs 2001, 2, 1070.
13. Goldstein, D. M.; Kuglstatter, A.; Lou, Y.; Soth, M. J. J. Med. Chem. 2010, 53,
2345.
a
% Turnover in human hepatic microsomes using a 30 min incubation period.
Passive permeability through a CACO-2 monolayer determined using 12.5 lM
b
test compound in the presence of 115 nM mannitol.
14. Representative patents include: (a) Angell, R. M.; Aston, N. M.; Bamborough, P.;
Bamford, M. J.; Cockerill, G. S.; Flack, S. S.; Laine, D. I.; Merrick, S. J.; Smith, K. J.;
Walker, A. L. PCT Int. Appl., WO2003032971, 2003; (c) Almansa, C.; Virgili, M.
PCT Int. Appl. WO2007000339, 2007; (c) Zhang, D.; Tasker, A.; Sham, K. K. C.;
Chakrabarti, P. P.; Falsey, R. J.; Herberich, B. J.; Pettus, L. H.; Rzasa, R. M. PCT Int.
Appl. WO2008045393, 2008; (d) Vidal, B.; Eastwood, P. R.; Gonzalez, J.; Esteve,
C. PCT Int. Appl. WO2008107125, 2008.
15. (a) Angell, R.; Aston, N. M.; Bamborough, P.; Buckton, J. B.; Cockerill, S.; de
Boeck, S. J.; Edwards, C. D.; Holmes, D. S.; Jones, K. L.; Laine, D. I.; Patel, S.;
Smee, P. A.; Smith, K. J.; Somers, D. O.; Walker, A. L. Bioorg. Med. Chem. Lett.
2008, 18, 4428; (b) Angell, R. M.; Angell, T. D.; Bamborough, P.; Bamford, M. J.;
Chung, C.; Cockerill, S. G.; Flack, S. S.; Jones, K. L.; Laine, D. I.; Longstaff, T.;
Ludbrook, S.; Pearson, R.; Smith, K. J.; Smee, P. A.; Somers, D. O.; Walker, A. L.
Bioorg. Med. Chem. Lett. 2008, 18, 4433.
C
Twenty-four hour agitation of 0.4 mg test compound in 0.4 mL aqueous buffer
at 37 °C followed by filtration and quantification by HPLC–UV.
Table 3
Pharmacokinetic profile of 16 in rat (iv 1 mg/kg)
Terminal
Cmax
Cl
AUC0–1
Vss
t½ (h)
(ng/mL)
(mL/min/kg)
(ng h/mL)
(L/kg)
5.3a
2002
6.6
2532
0.8
a
Refers to 8% of the dose during the second phase of elimination. Ninety per-
16. Wrobleski, S. T.; Lin, S.; Hynes, J.; Wu, H.; Pitt, S.; Shen, D. R.; Zhang, R.; Gillooly,
K. M.; Shuster, D. J.; McIntyre, K. W.; Doweyko, A. M.; Kish, K. F.; Tredup, J. A.;
Duke, G. J.; Sack, J. S.; McKinnon, M.; Dodd, J.; Barrish, J. C.; Schieven, G. L.;
Leftheris, K. Bioorg. Med. Chem. Lett. 2008, 18, 2739; (b) Hynes, J., Jr.; Dyckman,
A. J.; Lin, S.; Wrobleski, S. T.; Hong, Wu; Gillooly, K. M.; Kanner, S. B.; Lonial, H.;
Loo, D.; McIntyre, K. W.; Pitt, S.; Shen, D. R.; Shuster, D. J.; Yang, X.; Zhang, R.;
Behnia, K.; Zhang, H.; Marathe, P. H.; Doweyko, A. M.; Tokarski, J. S.; Sack, J. S.;
Pokross, M.; Kiefer, S. E.; Newitt, J. A.; Barrish, J. C.; Dodd, J.; Schieven, G. L.;
Leftheris, K. J. Med. Chem. 2008, 51, 4.
17. For further discussion on the ‘flip’ binding mode of unrelated series see: (a)
Fitzgerald, C. E.; Patel, S. B.; Becker, J. W.; Cameron, P. M.; Zaller, D.; Pikounis, V.
B.; O’Keefe, S. J.; Scapin, G. Nat. Struct. Biol. 2003, 10, 764; (b) Natarajan, S. R.;
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18. Kende, A. S.; Hodges, J. C. Synth. Commun. 1982, 12, 1.
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F.; Hayes, J. S. J. Med. Chem. 1987, 30, 824; (b) Di, M. A.; Foulon, L.; Garcia, G.;
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20. For full details of the biological assays used in this publication see:
Lumeras, W.; Caturla, F.; Vidal, L.; Esteve, C.; Balagué, C.; Orellana, A.;
Domínguez, M.; Roca, R.; Huerta, J. M.; Godessart, N.; Vidal, B. J. Med. Chem.
2009, 52, 5531.
centage of the dose is eliminated during the 1st phase of elimination with t½ < 1 h.
panels, the most significant findings being the inhibition of 5HT1B
(87% inhibition at 10 lM) in a panel of 50 receptors (Cerep, France)
and the inhibition of human c-Raf (IC50 = 200 nM) in a panel of 50
kinases (Upstate). No major concern arose when lead compound 16
was tested for inhibition of human cytochromes (IC50 data: 3A4,
2D6 > 25
lM; 2C19ꢁ25
l
M; 1A2, 2C9ꢁ10
lM).
The iv pharmacokinetic profile of compound 16 in the rat is pre-
sented in Table 3. Two elimination phases were seen: an initial
elimination phase with a short t½ < 1 h representing >90% of the
dose; and a terminal phase with t½ = 5.3 h representing the
remainder. Both a low clearance and low volume of distribution
were noted. Upon oral dosing at 10 mg/kg low levels of product
were detected (Cmax = 367 ng/mL) with a resulting bioavailability
of ꢁ10%. Factors such as first pass metabolism (high metabolism
in hepatic microsomes NADPH) and poor solubility (3
pH 7.4 = ꢁ8 M) are likely to contribute to the poor oral bioavail-
ability of 16.
In summary, structure-based design led to a new series of
potent and selective indolin-2-one p38 inhibitors. The subse-
lg/mL at
l
21. Coordinates and structure factors for the complex of p38a with 16 have been
deposited in the RCSB Protein Data Bank (access code 3RIN).
22. Eastwood, P. et al., unpublished results.
23. Citotoxicity assay: Trypsinized CHO cells (10,000 cells /well) were incubated in
96-well plates in a culture medium for 24 h at 37 °C with 5% CO2. The culture
medium was removed and the plates washed with dPBS (Biowhittaker BE-17-
512F or Sigma D8537). Hundred microliters of culture medium containing the
test compounds was added to the plate wells and incubation was continued for
a
quent optimization of this series will be the subject of further
publications.
24 h. The cells were then washed with 100 lL of dPBS and the reduction of the
References and notes
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