3
094
S. H. Mohd-Pahmi et al. / Bioorg. Med. Chem. Lett. 21 (2011) 3092–3094
Table 1
Kinetic data for inhibitors against pPAP and rkbPAP at pH 4.9
2. Denu, J. M.; Stuckey, J. A.; Saper, M. A.; Dixon, J. E. Cell 1996, 87, 361.
3. Mertz, P.; Yu, L.; Sikkink, R.; Rusnak, F. J. Biol. Chem. 1997, 272, 21296.
4.
Barford, D.; Das, A. K.; Egloff, M. P. Annu. Rev. Biophys. Biomol. Struct. 1998, 27,
Inhibitor
pPAP
rkbPAP
133.
5
6
.
.
Twitchett, M. B.; Sykes, A. G. Eur. J. Inorg. Chem. 1999, 2105.
Mueller, E. G.; Crowder, M. W.; Averill, B. A.; Knowles, J. R. J. Am. Chem. Soc.
1993, 115, 2974.
K
ic (mM)
K
iuc (mM)
K
ic (mM)
K
iuc (mM)
6
6
6
6
6
6
6
a
b
c
d
e
f
0.363 ± 0.165
0.116 ± 0.045
0.044 ± 0.013
0.010 ± 0.003
0.021 ± 0.007
0.008 ± 0.001
0.013 ± 0.003
1.439 ± 1.510
0.349 ± 0.225
0.238 ± 0.010
0.222 ± 0.109
0.195 ± 0.120
0.057 ± 0.024
0.005 ± 0.002
0.011 ± 0.004
0.031 ± 0.019
0.654 ± 0.240
0.446 ± 0.144
0.443 ± 0.198
0.102 ± 0.026
––
7. Klabunde, T.; Krebs, B. Struct. Bonding (Berlin) 1997, 89, 177.
8
.
Schenk, G.; Ge, Y.; Carrington, L. E.; Wynne, C. J.; Searle, I. R.; Carroll, B. J.;
Hamilton, S.; de Jersey, J. Arch. Biochem. Biophys. 1999, 370, 183.
Wilcox, D. E. Chem. Rev. 1996, 96, 2435.
––
––
––
––
––
9
.
1
1
1
1
1
1
1
1
0. Miti c´ , N.; Valizadeh, M.; Leung, E. W. W.; de Jersey, J.; Hamilton, S.; Hume, D.
A.; Cassady, A. I.; Schenk, G. Arch. Biochem. Biophys. 2005, 439, 154.
1. Schenk, G.; Elliott, T. W.; Leung, E.; Carrington, L. E.; Mitic, N.; Gahan, L. R.;
Guddat, L. W. B. M. C. Struct. Biol. 2008, 8, 6.
––
––
g
K
K
–
ic, Competitive inhibition constant.
iuc, Uncompetitive inhibition constant.
–, no significant uncompetitive inhibition.
2. McGeary, R. P.; Vella, P.; Mak, J. Y. W.; Guddat, L. W.; Schenk, G. Bioorg. Med.
Chem. Lett. 2009, 19, 163.
3. Zenger, S.; Hollberg, K.; Ljusberg, J.; Norgard, M.; Ek-Rylander, B.; Kiviranta, R.;
Andersson, G. Bone 2007, 41, 820.
4. Schwender, C. F.; Beers, S. A.; Malloy, E.; Demarest, K.; Minor, L.; Lau, K. H. W.
Bioorg. Med. Chem. Lett. 1995, 5, 1801.
5. Oddie, G. W.; Schenk, G.; Angel, N. Z.; Walsh, N.; Guddat, L. W.; de Jersey, J.;
Cassady, A. I.; Hamilton, S. E.; Hume, D. A. Bone 2000, 27, 575.
6. Leung, E.; Teixeira, M.; Guddat, L. W.; Miti c´ , N.; Schenk, G. Curr. Topics. Plant
Biol. 2007, 8, 21.
refluxing the corresponding carboxylic acids with thionyl chlo-
5
2
ride. The product amides were purified by flash column chroma-
tography to give 5a–g in good yields. The final synthetic step to
produce the newly designed inhibitors 6a–g required the cleavage
of the phosphonate ester bonds of 5a–g to give their corresponding
free phosphonic acids. This was achieved by heating 5a–g with a
mixture of sodium iodide and trimethylsilyl chloride in acetonitrile
7. Schenk, G.; Guddat, L. W.; Ge, Y.; Carrington, L. E.; Hume, D. A.; Hamilton, S.; de
Jersey, J. Gene 2000, 250, 117.
18. Mauck, K. F.; Clarke, B. L. Mayo Clin. Proc. 2006, 81, 662.
19. Boonen, S. Bone 2007, 40, S26.
12
at 40 °C giving the desired products in moderate yields.
20. Halleen, J. M.; Tiitinen, S. L.; Ylipahkala, H.; Fagerlund, K. M.; Väänänen, H. K.
The inhibitory effects of phosphonates 6a–g were tested against
Clin. Lab. 2006, 52, 499.
both pPAP and rkbPAP at pH 4.9.3
4,35
The results are listed in
21. Vella, P.; McGeary, R. P.; Gahan, L. R.; Schenk, G. Curr. Enzyme Inhib. 2010, 6,
18.
1
Table 1. The shorter-chain inhibitors 6a–b exhibited mixed mode
inhibition (competitive and uncompetitive) for pPAP, and inhibi-
tors 6a–d also showed this behaviour against rkbPAP. Such mixed
mode inhibition has also been reported for our previously reported
2
2
2. Reszka, A. A.; Rodan, G. A. Curr. Rheumatol. Rep. 2003, 5, 65.
3. Drake, M. T.; Clarke, B. L.; Khosla, S. Mayo Clin. Proc. 2008, 83, 1032.
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2
2
5. Martin, T. J.; Seeman, E. Best Pract. Res. Clin. Endocrinol. Metab. 2008, 22, 701.
6. Zaidi, M.; Moonga, B.; Moss, D. W.; Macintyre, I. Biochem. Biophys. Res. Commun.
1
2
inhibitors. For longer chain-length inhibitors, only competitive
inhibition was observed against both pPAP and rkbPAP. This
behaviour may reflect a stronger anchoring effect of the longer
alkyl chains into the groove adjacent to the active site of the en-
zyme, which would favour competitive inhibition. In agreement
with our modelling studies, the inhibitory potencies of these com-
pounds generally increased with increasing chain length, reaching
1
989, 159, 68.
27. Moonga, B. S.; Moss, D. W.; Patchell, A.; Zaidi, M. J. Physiol. (Lond.) 1990, 429,
9.
2
2
2
8. Wang, Y. L.; Andersson, G. Arch. Biochem. Biophys. 2007, 461, 85.
9. Lu, X. M.; Su, N.; Yang, J.; Huang, W.; Li, C.; Zhao, L.; He, Q. F.; Du, X. L.; Shen, Y.;
Chen, B.; Chen, L. Biochem. Biophys. Res. Commun. 2009, 390, 494.
0. Hasegawa, S.; Yonezawa, T.; Ahn, J. Y.; Cha, B. Y.; Teruya, T.; Takami, M.;
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1. Bonnick, S. L.; Shulman, L. Am. J. Med. 2006, 119, 25S.
32. Hayman, A. R.; Jones, S. J.; Boyde, A.; Foster, D.; Colledge, W. H.; Carlton, M. B.;
Evans, M. J.; Cox, T. M. Development 1996, 122, 3151.
3. Angel, N. Z.; Walsh, N.; Forwood, M. R.; Ostrowski, M. C.; Cassady, A. I.; Hume,
3
3
a maximum Kic of 8
lM for 6f against pPAP and 5
l
M for 6e against
rkbPAP.
3
3
3
3
In conclusion, new inhibitors of pPAP and rkbPAP have been
identified using computer-aided design, and these have been
shown to exhibit potent inhibitory activities against these en-
D. A. J. Bone Miner. Res. 2000, 15, 103.
4. Valizadeh, M.; Schenk, G.; Nash, K.; Oddie, G. W.; Guddat, L. W.; Hume, D. A.; de
Jersey, J.; Burke, T. R.; Hamilton, S. Arch. Biochem. Biophys. 2004, 424, 154.
5. Elliott, T. W.; Miti c´ , N.; Gahan, L. R.; Guddat, L. W.; Schenk, G. J. Braz. Chem. Soc.
zymes. Compound 6e (Kic, 5
potent inhibitor of rkbPAP (4
lM) is comparable to that of the most
2006, 17, 1558.
12
lM) and compound 6f (Kic, 8 lM) is
6. Pinkse, M. W. H.; Merkx, M.; Averill, B. A. Biochemistry 1999, 38, 9926.
the most potent inhibitor of pPAP yet described. In addition, it has
also been shown that predicted binding affinities calculated using
computational modelling correlate well with measured inhibition
constants for this class of inhibitor.
37. Wang, X.; Ho, R. Y. N.; Whiting, A. K.; Que, L., Jr. J. Am. Chem. Soc. 1999, 121,
235.
9
3
3
8. Crans, D. C.; Simone, C. M.; Holz, R. C.; Que, L. Biochemistry 1992, 31, 11731.
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4
4
4
4
1. Marshall, K.; Nash, K.; Haussman, G.; Cassady, I.; Hume, D.; de Jersey, J.;
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Acknowledgments
2. Sträter, N.; Jasper, B.; Scholte, M.; Krebs, B.; Duff, A. P.; Langley, D. B.; Han, R.;
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We thank Dr. Tri Le for assistance with NMR studies and
Mr. Graham MacFarlane for MS measurements. Mr. Peter Vella
and Mr. Daniel Feder are thanked for helpful discussions. This work
was funded by a grant from the Australian Research Council
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4
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