H. Y. Lo et al. / Bioorg. Med. Chem. Lett. 21 (2011) 4533–4539
4539
Table 4
Miyazaki, M. Life Sci. 2002, 71, 437; (c) Jin, D.; Takai, S.; Yamada, M.; Sakaguchi,
M.; Kamoshita, K.; Ishida, K.; Sukenaga, Y.; Miyazaki, M. Cardiovasc. Res. 2003,
60, 413; (d) Palaniyandi, S. S.; Nagai, Y.; Watanbe, K.; Ma, M.; Veeraveedu, P. T.;
Prakash, P.; Kamal, F. A.; Abe, Y.; Yamaguchi, K.; Tachikawa, H.; Kodama, M.;
Aizawa, Y. Exp. Biol. Med. 2007, 232, 1213.
Molecular calculation of oxidative energy of P1
Compounds
HOMOa (kcal/mol)
LUMOa (kcal/mol)
D
Ea (kcal/mol)
1
31
41
36
21
32
ꢀ133.6
ꢀ121.7
ꢀ118.8
ꢀ125.7
ꢀ129.8
ꢀ141.9
ꢀ20.08
ꢀ10.04
ꢀ9.933
ꢀ12.65
ꢀ34.34
ꢀ23.38
173.0
159.6
155.0
160.9
167.4
215.4
8. Aoyama, Y. Expert Opin. Ther. Patents 2001, 11, 1423.
10. Chymase was assayed in a 15
human chymase and 100 nM rhodamine 110, bis-(succinoyl-
-prolyl- -phenylalanylamide) (American Peptide) in buffer containing
20 mM Tris HCl pH 8.0, 50 mM NaCl, 100 M TCEP, 0.01% CHAPS, and 1%
l
L volume containing 500 pM recombinant
L
-alanyl- -alanyl-
L
L
L
a
l
a
Molecular calculation using quantum chemistry DFT/B3LYP method.
DMSO for 1 h at 28 °C/80% humidity, and fluorescence was read at 485 nm
excitation and 530 nm emission.
11. The coordinates of co-crystal structure of 2 with human chymase are deposited
in the RCSB protein data bank (RCSD ID: RCSB065611; PDB ID: 3S0N).
12. To assess reactive metabolite formation, compound 1 (50 lM) was added to
most potent. However, only the benzoisothiazole P1 prevents the
formation of a GSH conjugate. The calculated oxygenation energies
of the new P1 analogs agree with the experimental results. The
progress of this new class of chymase inhibitor toward a new drug
candidate will be reported in the near future.
human liver microsomes (1 mg/mL) in phosphate buffer (100 mM, pH 7.4).
Pooled human liver microsomes were purchased from BD Biosciences (San
Jose, CA). After preincubation at 37 °C for 3 min, the reaction was initiated by
the addition of NADPH (2.5 mM). The final incubation volume was 1 mL.
Samples without compound or NADPH were used as negative controls. After
incubation for 2 h, samples were quenched with 2 mL of acetonitrile. Samples
were centrifuged at 2095ꢂg for 5 min and the supernatant was transferred to a
Acknowledgment
glass tube and the solvent was evaporated using
(Hopkinton, MA). Samples were redissolved in 500
a
Caliper TurboVap LV
l
L water/acetonitrile (1:1,
The authors would like to thank Dr. Anil Padyana for the depo-
sition of X-ray crystal coordinates.
v:v) and analyzed by mass spectrometry for the detection of glutathione
adducts. A glutathione adduct of compound 1 was detected after analysis of the
microsomal incubate.
13. (a) Uetrecht, J. P. Curr. Drug Metab. 2000, 1, 133; (b) Zhou, S.; Chan, E.; Duan,
W.; Huang, M.; Chen, Y.-Z. Drug Metab. Rev. 2005, 1, 41; (c) Walgren, J. L.;
Mitchell, M. D.; Thompson, D. C. Crit. Rev. Toxicol. 2005, 35, 325.
Reference and notes
1. Bacani, C.; Frishman, W. H. Cardiol. Rev. 2006, 14, 187.
14. Cathepsin G was assayed as described for chymase in 15 lL containing 4 nM
2. (a) McEuen, A. R.; Sharama, B.; Walls, A. F. Biochim. Biophys. Acta 1995, 1267,
115; (b) Galli, S. J.; Wershil, B. K. Exp. Dermatol. 1995, 4, 240; (c) Reiling, K. K.;
Krucinski, J.; Miercke, L. J., et al Biochemistry 2003, 42, 2616.
3. (a) Nishimoto, M.; Takai, S.; Kim, S.; Jin, D.; Yuda, A.; Sakaguchi, M.; Yamada,
M.; Sawada, Y.; Kondo, K.; Asada, K.; Iwao, H.; Sasaki, S.; Miyazaki, M.
Circulation 2001, 104, 1274; (b) Nishimoto, M.; Takai, S.; Sawada, Y.; Yuda, A.;
Kondo, K.; Yamada, M.; Jin, D.; Sakaguchi, M.; Asada, K.; Sasaki, S.; Miyazaki, M.
J. Thorac. Cardiovasc. Surg. 2001, 121, 729.
4. (a) Takai, S.; Miyazaki, M. Curr. Vasc. Pharmacol. 2003, 1, 217; (b) Doggrell, S. A.;
Wanstall, J. C. Cardiovasc. Res. 2004, 61, 653.
5. Taipale, J.; Lohi, J.; Saarinen, J.; Kovanen, P. T.; Keski-Oja, J. J. Biol. Chem. 1995,
270, 4689.
cathepsin G (Athens Research and Technology) and 300 nM chymase substrate
in a buffer containing 1ꢂ phosphate-buffered saline, 10 mM HEPES pH 7.4,
100 lM TCEP, 0.01% CHAPS, and 1% DMSO.
15. Dalpozzo, R.; Bartoli, G. Curr. Org. Chem. 2005, 9, 163.
16. (a) Kubow, S.; Janzen, E. G.; Bray, T. M. J. Biol. Chem. 1984, 259, 4447; (b)
Nocerini, M. R.; Yost, G. S.; Carlson, J. R.; Liberato, D. J.; Breeze, R. G. Drug Metab.
Dispos. 1985, 13, 690; (c) Thornton-Manning, J.; Appleton, M. L.; Gonzalez, F. J.;
Yost, G. S. J. Pharmacol. Exp. Ther. 1996, 276, 21; (d) Zhang, K. E.; Kari, P. H.;
Davis, M. R.; Doss, G.; Baillie, T. A.; Vyas, K. P. Drug Metab. Dispos. 2000, 28, 633.
17. Unpublished results for the acid side chain modification.
18. Energy calculations were carried out using quantum chemistry DFT/B3LYP
method. The basis-set function used in the calculation was 6-31⁄⁄G++. The
geometry of each group was optimized by the DFT method and the energy
value is for the fully optimized molecule.
6. Kofford, M. W.; Schwartz, L. B.; Schechter, N. M., et al J. Biol. Chem. 1997, 272,
7127.
7. (a) Jin, D.; Takai, S.; Yamada, M.; Sakaguchi, M.; Yao, Y.; Miyazaki, M. Jpn. J.
Pharmacol. 2001, 86, 203; (b) Jin, D.; Takai, S.; Yamada, M.; Sakaguchi, M.;