Daniel Burkett
Marquette
Development of Small Molecule Inhibitors of Pyruvate Carboxylase
Pyruvate Carboxylase (PC), a major anaplerotic enzyme, catalyzes the MgATP and bicarbonate-dependent carboxylation of pyruvate to oxaloacetate to replenish depleted citric acid cycle intermediates(1). Abnormal PC activity has been linked to microbial virulence2-3, Type-II diabetes (4-6), and tumor cell proliferation (7-9). Studies have shown therapeutic benefits of knocking down or inhibiting PC, making this enzyme an attractive target for small molecule inhibition (10). Based on the biological evaluation of simple metal chelators, biotin analogs, and a series of generated α-keto acids, the incorporation of this α-keto moiety appears crucial to inhibition. Additionally, the α-keto acid’s ability to exist as the enol tautomer correlates to it’s potency in inhibiting PC. 2-Oxo-3-(quinolin-2-yl)propanoic acid was found to be the most potent inhibitor (S. aureus PC IC50 = 5.7 ± 1.0 μM) and this is attributed to this compound’s ability to form an intramolecular hydrogen bond, stabilizing the enol tautomer. Progress toward the development of a potent, selective small molecule inhibitor of PC will be presented.
References
1. Lietzan, A.D.; Lin, Y.; St. Maurice, M. Arch. Biochem. Biophys. 2014, 562, 70-79.
2. Schar, J.; Stoll, R.; Schauer, K.; Loeffler, D.I.; Eylert, E.; Joseph, B.; Eisenreich, W.; Fuchs, T.M.; Goebel, W. J. Bacteriol. 2010, 192, 1774-1784.
3. Kukavica-Ibrulj, I.; Sanschagrin, F.; Peterson, A.; Whiteley, M.; Boyle, B.; Mackay, J.; Levesque, R.C. Microbiology, 2008, 154, 2106-2118.
4. Han, J.; Liu, Y.Q. J. Endocrinol. 2010, 204, 143-152.
5. MacDonald, M.J.; Longacre, M.J.; Langberg, E.C.; Tibell, A.; Kendrick, M.A.; Fukao, T.; Ostenson, C.G. Diabetologia, 2009, 52, 1087-1091.
6. Hasan, N.M.; Longacre, M.J.; Stoker, S.W.; Boonsaen, T.; Jitrapakdee, S.; Kendrick, M.A.; Wallace, J.C.; MacDonald, M.J. J. Biol. Chem. 2008, 283, 28048-28059.
7. Forbes, N.S.; Meadows, A.L.; Clark, D.S.; Blanch, H.W. Metabol. Eng., 2006, 8, 639-652.
8. Liu, K.J.; Kleps, R.; Henderson, T.; Nyhus, L. Biochem. Biophys. Res. Comm., 1991, 179, 366-371.
9. Fan, T.W.M.; Lane, A.N.; Higashi, R.M. Mol. Cancer, 2009, 8-41.
10. Zeczycki, T.N.; St. Maurice, M.; Attwood, P.V. Open Enzyme Inhib. J. 2010, 3, 8-26.
Pyruvate Carboxylase (PC), a major anaplerotic enzyme, catalyzes the MgATP and bicarbonate-dependent carboxylation of pyruvate to oxaloacetate to replenish depleted citric acid cycle intermediates(1). Abnormal PC activity has been linked to microbial virulence2-3, Type-II diabetes (4-6), and tumor cell proliferation (7-9). Studies have shown therapeutic benefits of knocking down or inhibiting PC, making this enzyme an attractive target for small molecule inhibition (10). Based on the biological evaluation of simple metal chelators, biotin analogs, and a series of generated α-keto acids, the incorporation of this α-keto moiety appears crucial to inhibition. Additionally, the α-keto acid’s ability to exist as the enol tautomer correlates to it’s potency in inhibiting PC. 2-Oxo-3-(quinolin-2-yl)propanoic acid was found to be the most potent inhibitor (S. aureus PC IC50 = 5.7 ± 1.0 μM) and this is attributed to this compound’s ability to form an intramolecular hydrogen bond, stabilizing the enol tautomer. Progress toward the development of a potent, selective small molecule inhibitor of PC will be presented.
References
1. Lietzan, A.D.; Lin, Y.; St. Maurice, M. Arch. Biochem. Biophys. 2014, 562, 70-79.
2. Schar, J.; Stoll, R.; Schauer, K.; Loeffler, D.I.; Eylert, E.; Joseph, B.; Eisenreich, W.; Fuchs, T.M.; Goebel, W. J. Bacteriol. 2010, 192, 1774-1784.
3. Kukavica-Ibrulj, I.; Sanschagrin, F.; Peterson, A.; Whiteley, M.; Boyle, B.; Mackay, J.; Levesque, R.C. Microbiology, 2008, 154, 2106-2118.
4. Han, J.; Liu, Y.Q. J. Endocrinol. 2010, 204, 143-152.
5. MacDonald, M.J.; Longacre, M.J.; Langberg, E.C.; Tibell, A.; Kendrick, M.A.; Fukao, T.; Ostenson, C.G. Diabetologia, 2009, 52, 1087-1091.
6. Hasan, N.M.; Longacre, M.J.; Stoker, S.W.; Boonsaen, T.; Jitrapakdee, S.; Kendrick, M.A.; Wallace, J.C.; MacDonald, M.J. J. Biol. Chem. 2008, 283, 28048-28059.
7. Forbes, N.S.; Meadows, A.L.; Clark, D.S.; Blanch, H.W. Metabol. Eng., 2006, 8, 639-652.
8. Liu, K.J.; Kleps, R.; Henderson, T.; Nyhus, L. Biochem. Biophys. Res. Comm., 1991, 179, 366-371.
9. Fan, T.W.M.; Lane, A.N.; Higashi, R.M. Mol. Cancer, 2009, 8-41.
10. Zeczycki, T.N.; St. Maurice, M.; Attwood, P.V. Open Enzyme Inhib. J. 2010, 3, 8-26.