Human fatty acid synthase (hFAS) is a complex, multifunctional enzyme that is solely responsible for the de novo synthesis of long chain fatty acids. hFAS is highly expressed in a number of cancers, with low expression observed in most normal tissues. Although normal tissues tend to obtain fatty acids from the diet, tumor tissues rely on de novo fatty acid synthesis, making hFAS an attractive metabolic target for the treatment of cancer. We describe here the identification of GSK2194069, a potent and specific inhibitor of the β-ketoacyl reductase (KR) activity of hFAS; the characterization of its enzymatic and cellular mechanism of action; and its inhibition of human tumor cell growth. We also present the design of a new protein construct suitable for crystallography, which resulted in what is to our knowledge the first co-crystal structure of the human KR domain and includes a bound inhibitor.
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We are grateful to C. Aquino for designing and first preparing GSK2194069 and to A. Chaudhari, J. Mack and K. Erhard for preparing additional quantities. We thank Y. Shen for construct engineering, J. Christie for virus generation and G. Barrett for large-scale cell culture. We thank B. Rodríguez, M. Jesús Vázquez and R. Liu for the high-throughput screening data; K. McKenzie and J. Gross for the initial FAS inhibition data for GSK2194069; and G. Waitt for the LC/MS/MS analyses in the M-CoA experiments. We also thank G. Graczyk-Millbrandt and M. Wang for final structure confirmation by NMR. Support of this program by R. Wooster is gratefully acknowledged. Use of the Advanced Photon Source was supported by the US Department of Energy, Office of Science, Office of Basic Energy Sciences, under contract no. DE-AC02-06CH11357. Use of the Life Sciences Collaborative Access Team (LS-CAT) Sector 21 was supported by the Michigan Economic Development Corporation and the Michigan Technology Tri-Corridor (grant no. 085P1000817).
All of the authors are employees of GlaxoSmithKline, and all are or have been shareholders of GlaxoSmithKline.
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Hardwicke, M., Rendina, A., Williams, S. et al. A human fatty acid synthase inhibitor binds β-ketoacyl reductase in the keto-substrate site. Nat Chem Biol 10, 774–779 (2014). https://doi.org/10.1038/nchembio.1603
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