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Figure 2. Inhibition of tissue TG accumulation in vivo (⁄P <0.05 versus HFD control. ⁄⁄P <0.01 versus HFD control).
organic phase was removed. To remove [1À14C] oleic acid completely, the
aqueous phase was washed by heptane again. The aqueous phase was added
Pico-Fluor 40 scintillation cocktail (PerkinElmer) and the radioactivity was
measured by LSC-6100 scintillation counter (Aloka). The radioactivity of FLAG-
tagged luciferase protein instead of FATP1 protein was deducted as
background. The IC50 values represent the average of at least n = 2.
5. The homology between human and mouse FATP1 is calculated to be 89% from NCBI
reference sequence: NP_940982.1 (human FATP1), NP_036107.1 (mouse FATP1).
6. Mouse microsomal stability was evaluated by measuring the remaining (%) of
the test compound after 0.5-h incubation with mouse liver microsome.
7. (a) Chordia, M.; Murphree, L. J.; Macdonald, T. L.; Linden, J.; Olsson, R. A. Bioorg.
Med. Chem. Lett. 2002, 12, 1563; (b) Lang, M.; Seifert, M. H.-J.; Wolf, K. K.;
Aschenbrenner, A.; Baumgartner, R.; Wieber, T.; Trentinaglia, V.; Blisse, M.;
Vitt, D.; Tajima, N.; Yamashita, T.; Noda, H. Bioorg. Med. Chem. Lett. 2011, 21,
5417.
8. For the synthesis of 1-(5-fluoropyridine-2-yl)piperazine hydrochloride, see: Slassi,
A.; Isac, M.; Arora, J.; Brown, D. PCT Int. Appl. 2007, WO 2007087135.
9. (a) Almirante, L.; Mugaini, A.; Toma, D. N.; Gamba, A.; Murmann, W. J. Med.
Chem. 1970, 13, 1048; (b) Hlasta, J. D. Tetrahedron Lett. 1990, 31, 5833; (c)
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Flanagan, J. U.; Hay, M. P.; Chan, D. A.; Lai, E. W.; Nguyen, P.; Giaccia, A. J. Bioorg.
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Holloway et al. reported that FATP1 overexpression in mice muscle
did not cause muscular TG accumulation or whole body insulin
resistance.10 Taken together, FATP1 inhibition alone may not have
enough of an impact on fatty acid transport and/or metabolism to
reduce tissue TG contents.
References and notes
1. (a) Schaffer, J. E.; Lodish, H. F. Cell 1994, 79, 427; (b) Abumrad, N.; Coburn, C.;
Ibrahimi, A. Biochem. Biophys. Acta 1999, 1441, 4; (c) Bonen, A.; Miskovic, D.;
Kiens, B. Can. J. Appl. Physiol. 1999, 24, 515; (d) Kazantzis, M.; Stahl, A. Biochem.
Biophys. Acta 2012, 1821, 852.
2. Kim, J. K.; Gimeno, R. E.; Higashimori, T.; Kim, H.-J.; Choil, H.; Punreddy, S.;
Mozell, R. L.; Tan, G.; Stricker-Krongrad, A.; Hirsch, D. J.; Fillmore, J. J.; Liu, Z.-X.;
Dong, J.; Cline, G.; Stahl, A.; Lodish, H. F.; Shulman, G. I. J. Clin. Invest. 2004, 113,
756.
3. Matsufuji, T.; Ikeda, M.; Naito, A.; Kanda, S.; Izumi, M.; Harada, J.; Takakusa, H.;
Hirouchi, M.; Shinozuka, T. Bioorg. Med. Chem. Lett. 2012, 22, 5067.
4. The inhibition of human or mouse FATP1 was evaluated by cell-free ACS assay:
FLAG-tag fused human or mouse FATP1 protein was incubated with 10 lM
[1-14C] oleic acid and test compound in 100 mM Tris–HCl (pH 7.4), 30 mM
NaCl, 0.5 mM ATP, 5 mM MgCl2, 0.5 mM coenzyme A, 0.05% Triton-X 100,
2 mM dithiothreitol at 37 °C for 20 min. After the reaction, [1-14C] oleoyl-CoA
was extracted by successive additions of fivefold volume of 2-propanol/
heptane/H2SO4 (40:10:1, v/v/v), threefold volume of heptane, and twofold
volume of water. Phases were then separated by centrifugation, and the
10. Holloway, G. P.; Chou, C. J.; Lally, J.; Stellingwerff, T.; Maher, A. C.; Gavrilova, O.;
Haluzik, M.; Alkhateeb, H.; Reitman, M. L.; Bonen, A. Diabetologia 2011, 54,
1457.