H. P. Beck et al. / Bioorg. Med. Chem. Lett. 21 (2011) 2752–2755
2755
7. PBS = phosphate buffer saline; for experimental details see Tan, H.; Semin, D.;
Wacker, M.; Cheetham, J. J. Assoc. Lab. Autom. 2005, 10, 364.
8. For chiral separation procedure see Ref. 5.
in PBS. We characterized these analogs in rat in vivo PK experi-
ments (Table 3). Acid 20 exhibited high clearance of 5.7 L/h/kg.
However, 23 demonstrated low clearance (0.32 L/h/kg) and had
modest oral bioavailability (F 54%).
In conclusion, by blocking the metabolic soft-spot we were able
to substantially improve the PK properties of the chromenotriazol-
opyrimidine series of MDM2–p53 inhibitors while maintaining
both the biochemical and cellular potencies. Further in vivo phar-
macodynamic experiments of this series will be reported in due
course.
9. Compound (1 lM) was incubated with liver microsomes (0.25 mg/mL) and
NADPH (1 mM) in PBS for 30 min.
10. For example of incorporation of water-solubilizing moieties to improve
properties of MDM2 inhibitors see: Leonard, K.; Marugan, J. J.; Raboisson, P.;
Calvo, R.; Gushue, J. M.; Koblish, H. K.; Lattanze, J.; Zhao, S.; Cummings, M. D.;
Player, M. R.; Maroney, A. C.; Lu, T. Bioorg. Med. Chem. Lett. 2006, 16, 3463.
11. For hypothesized mechanism of epimerization see Ref. 5.
12. Stronger bases such as LiH resulted in O-alkylation.
N
N
N
N
O
S
Acknowledgements
The authors thank Dr. Michael Bartberger for helpful discus-
sions and Dr. Manuel Ventura for chiral separations.
Br
Br
References and notes
13. Both m-CPBA and Dess-Martin periodinate failed to provide any acid 11.
14. All biologically active compounds were characterized by 1H NMR, and their
purity was determined to be >95% by reverse phase HPLC; 1H NMR (Bruker
either 400 or 500 MHz), HPLC (A: 0.1% TFA in H2O, B: 0.1% TFA in CH3CN, 10–
100% B in 15 min.
15. Kane, S. A.; Fleener, C. A.; Zhang, Y. S.; Davis, L. J.; Musselman, A. L.; Huang, P. S.
Anal. Biochem. 2000, 278, 29.
16. mRNA p21 levels in SJSA-1 cells with 10% FBS were measured using Taqman;
see Ref. 5 for experimental details; the standard deviation for benchmark
compound 1.0, (n = 34).
1. (a) Vazquez, A.; Bond, E. E.; Levine, A. J.; Bond, G. L. Nat. Rev. Drug Disc. 2008, 7,
979; (b) Fridman, J. S.; Lowe, S. W. Oncogene 2003, 22, 9030.
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M. H. G.; Jones, S. N.; Vousden, K. H. Nature 1997, 387, 299; (c) Midgley, C. A.;
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17. 1H NMR of 19 (400 MHz, MeOH) d ppm 4.79 (s, 1H) 4.84 (s, 1H) 5.62 (s, 1H)
6.06 (s, 1H) 6.75–6.84 (m, 3H) 6.96–7.03 (m, 2H) 7.06–7.12 (m, 3H) 7.12–7.18
(m, 2H) 7.20–7.28 (m, 1H) 7.51 (dd, J = 7.83, 1.57 Hz, 1H) 7.70 (s, 1H).
18. 1H NMR of 21 (400 MHz, chloroform-d) d ppm 1.56–1.67 (m, 4H) 2.37 (t,
J = 6.65 Hz, 2H) 4.21 (t, J = 7.53 Hz, 2H) 5.50 (s, 1H) 5.84 (s, 1H) 6.79 (dq,
J = 8.90, 4.47 Hz, 4H) 6.98 (dd, J = 8.22, 1.17 Hz, 1H) 7.13–7.20 (m, 3H) 7.22–
7.26 (m, 2H) 7.34 (ddd, J = 8.22, 7.34, 1.47 Hz, 1H) 7.48 (dd, J = 7.83, 1.57 Hz,
1H) 8.03 (br s, 1H).
3. (a) Rayburn, E.; Zhang, R.; He, J.; Wang, H. Curr. Cancer Drug Targets 2005, 5, 27;
(b) Polsky, D.; Bastin, B. C.; Hazan, C.; Melzer, K.; Pack, J.; Houghton, A.; Busam,
K.; Cordon-Cardo, C.; Osman, I. Cancer Res. 2001, 61, 7642; (c) See 2d and 2e.
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2009, 49, 223; (b) Hu, C.-Q.; Hu, Y.-Z. Curr. Med. Chem. 2008, 15, 1720; (c) Patel,
S.; Player, M. R. Expert Opin. Invest. Drugs 2008, 17, 1865.
5. Allen, J. G.; Bourbeau, M. P.; Wohlhieter, G. E.; Bartberger, M. D.; Michelsen, K.;
Hungate, R.; Gadwood, R. C.; Gaston, R. D.; Evans, B.; Mann, L. W.; Matison, M.
E.; Schneider, S.; Huang, X.; Yu, D.; Andrews, P. S.; Reichelt, A.; Long, A. M.;
Yakowec, P.; Yang, E. Y.; Lee, T. A.; Oliner, J. D. J. Med. Chem. 2009, 52, 7044.
6. Iv 2.0 mg/kg dose in DMSO; po 5.0 mg/kg dose in 2% HPMC, 1% Tween 80, pH
2.2 witsh HCl.
19. For 20: OD-H column with SFC [24 g/min MeOH/0.2% diethylamine + 46 g/min
CO2]; for 23: separated at ester 18 250 Â 30 mm OD-H column with SFC [20 g/
min MeOH/0.2% diethylamine + 60 g/min CO2].