Discovery of small molecule isozyme non-specific inhibitors of mammalian acetyl-CoA carboxylase 1 and 2

Bioorganic and Medicinal Chemistry Letters

Volumn 20, Issue 7, 2010, Pages 2383-2388

  Corbett, Jeffrey W ^a   Freeman Cook, Kevin D ^a   Elliott, Richard ^a   Vajdos, Felix ^a   Rajamohan, Francis ^a   Kohls, Darcy ^a   Marr, Eric ^a   Zhang, Hailong ^b   Tong, Liang ^b   Tu, Meihua ^a   Murdande, Sharad ^a   Doran, Shawn D ^a   Houser, Janet A ^a   Song, Wei ^a   Jones, Christopher J ^a   Coffey, Steven B ^a   Buzon, Leanne ^a   Minich, Martha L ^a   Dirico, Kenneth J ^a   Tapley, Susan ^a   Kirk McPherson, R ^a   Sugarman, Eliot ^a   James Harwood Jr , H ^a   Esler, William ^a   more..

^a PFIZER INC   (United States)

^b Och Spine at New York Presbyterian Hospitals   (United States)

Author keywords

ACC; ACC1; ACC2; Acetyl CoA carboxylase; Diabetes; Enzyme inhibitor; Metabolic syndrome; Spirochromanone; Structure based drug discovery

Indexed keywords

ACETYL COENZYME A CARBOXYLASE; ACETYL COENZYME A CARBOXYLASE 1; ACETYL COENZYME A CARBOXYLASE 2; ANTHRACENE DERIVATIVE; LIGASE INHIBITOR; QUINOLINE DERIVATIVE; SPIROCHROMANONE DERIVATIVE; UNCLASSIFIED DRUG;

ANIMAL EXPERIMENT; ARTICLE; DOG; DRUG ABSORPTION; DRUG BINDING; DRUG BIOAVAILABILITY; DRUG CLEARANCE; DRUG HALF LIFE; DRUG SCREENING; DRUG STRUCTURE; DRUG SYNTHESIS; FATTY ACID OXIDATION; FATTY ACID SYNTHESIS; HYDROGEN BOND; MAXIMUM PLASMA CONCENTRATION; NONHUMAN; RAT; STRUCTURE ACTIVITY RELATION;

ACETYL-COA CARBOXYLASE; ANIMALS; ENZYME INHIBITORS; HUMANS; ISOENZYMES; MODELS, MOLECULAR; RATS; SMALL MOLECULE LIBRARIES; STRUCTURE-ACTIVITY RELATIONSHIP;

MAMMALIA; RATTUS;

EID: 77949489998     PISSN: 0960894X     EISSN: None     Source Type: Journal    
DOI: 10.1016/j.bmcl.2009.04.091     Document Type: Article

References (17)

1. Data was obtained from the World Health Organization website: http://www.who.int/diabetes/en/.
2. From WHO's website: http://www.who.int/mediacentre/factsheets/fs311/en/index.html.
3. For instance, see the guidelines for management of hyperlipidemia in children and adolescents put forward by the Kansas City Quality Improvement Consortium: http://kcqic.org/wp-content/uploads/2008/05/2007%20KCQIC%20Hyperlipidemia%20Guideline%20%20Children%20and%20Adolescents.pdf.
4. For recent reviews discussing acetyl-CoA carboxylase as a potential drug target, see:. Harwood Jr. H.J. Expert Opin. Cardiovasc. Renal 9 (2005) 267
5. Tong L., and Harwood Jr. H.J. J. Cell. Biochem. 99 (2006) 1476
6. Corbett J.W., and Harwood Jr. H.J. Recent Patents Cardiovasc. Drug Disc. 2 (2007) 162
7. Polakis S.E., Guchhait R.B., Zwergel E.S., Lane M.D., and Cooper T.G. J. Biol. Chem. 249 (1974) 6657
8. Harwood Jr. H.J., Petras S.F., Shelly L.D., Zaccaro L.M., Perry D.A., Makowski M.R., Hargrove D.M., Martin K.A., Tracey W.R., Chapman J.G., Magee W.P., Dalvie D.K., Soliman V.F., Martin W.H., Mularski C.J., and Eisenbeis S.A. J. Biol. Chem. 278 (2003) 37099
9. For the yeast CT-domain co-crystal structure with CP-640186, see:. Zhang H., Tweel B., Li J., and Tong L. Structure 12 (2004) 1683
10. Vajdos, F.; Rajamohan, F.; Marr, E.; Corbett, J. W., in preparation.
11. Hopkins A.L., Groom C.R., and Alex A. Drug Discovery Today 9 (2004) 430
12. Rat ACC1 was obtained from rat liver based upon standard procedures such as those described by. Thampy K.G., and Wakil S.J. J. Biol. Chem. 260 (1985) 6318 The assay was conducted as described in Ref. 6
13. Subsequent to the initiation of our work, the following patent applications published: (a) JP 2005119987; (b) WO 2007011809; (c) WO 2007011811.
14. Ascension numbers for the crystal structures are: compound 2 = 3h0j and compound 7 = 3h0s.
15. Abu-Elheiga L., Jayakumar A., Baldini A., Chirala S., and Wakil S. Proc. Natl. Acad. Sci. U.S.A. 92 (1995) 4011
16. Human ACC2 inhibition is measured using purified recombinant human ACC2 (hACC2). A full length Cytomax clone of hACC2 was purchased from Cambridge Bioscience Limited and was sequenced and subcloned into PCDNA5 FRT TO-TOPO (Invitrogen, Carlsbad, CA). The hACC2 was expressed in CHO cells by tetracycline induction and harvested in 5 L of DMEM/F12 with glutamine, biotin, hygromycin and blasticidin with 1 μg/mL tetracycline. The conditioned medium containing hACC2 was then applied to a Softlink Soft Release Avidin column (Promega, Madison, WI) and eluted with 5 mM biotin. hACC2 (4 mg) was eluted at a concentration of 0.05 mg/mL with an estimated purity of 95%. The purified hACC2 was dialyzed in 50 mM Tris, 200 mM NaCl, 4 mM DTT, 2 mM EDTA, and 5% glycerol. The pooled protein was frozen and stored at -80 °C, with no loss of activity upon thawing. For measurement of hACC2 activity and assessment of hACC2 inhibition, test compounds are dissolved in DMSO and added to the hACC2 enzyme as a 5× stock with a final DMSO concentration of 1%. rhACC2 was assayed in a Costar #3767 (Costar, Cambridge, MA) 384-well plate using the Transcreener ADP detection FP assay kit (Bellbrook Labs, Madison, WI) using the manufacturers' conditions for a 50 μM ATP reaction. The final conditions for the assay are 50 mM HEPES, pH 7.5, 5 mM MgCl2, 5 mM tripotassium citrate, 2 mM DTT, 0.5 mg/mL BSA, 30 μM acetyl-CoA, 50 μM ATP, and 8 mM KHCO3. Typically, a 10 μL reaction is run for 1 h at room temperature and 10 μL of Transcreener stop and detect buffer is added and incubated for an additional 1 h. The data is acquired on an Envision Fluorescence reader (Perkin Elmer) using a 620 excitation Cy5 FP general dual mirror, 620 excitation Cy5 FP filter, 688 emission (S) and a 688 (P) emission filter.
17. 6): 12.9 (s, 1H), 8.1 (d, J = 5.8 Hz, 1H), 7.6 (s, 1H), 7.2 (s, 1H), 6.7 (d, J = 5.9 Hz), 5.4 (m, 1H), 2.8 (s, 2H), 1.3 (d, J = 6.2 Hz, 6H).