A small-molecule AdipoR agonist for type 2 diabetes and short life in obesity (original) (raw)

References

  1. Gesta, S., Tseng, Y. H. & Kahn, C. R. Developmental origin of fat: tracking obesity to its source. Cell 131, 242–256 (2007)
    Article CAS Google Scholar
  2. Olefsky, J. M. & Glass, C. K. Macrophages, inflammation, and insulin resistance. Annu. Rev. Physiol. 72, 219–246 (2010)
    Article CAS Google Scholar
  3. Osler, M. E. & Zierath, J. R. Adenosine 5′-monophosphate-activated protein kinase regulation of fatty acid oxidation in skeletal muscle. Endocrinology 149, 935–941 (2008)
    Article CAS Google Scholar
  4. LeRoith, D. & Accili, D. Mechanisms of disease: using genetically altered mice to study concepts of type 2 diabetes. Nature Clin. Pract. Endocrinol. Metab. 4, 164–172 (2008)
    Article CAS Google Scholar
  5. Scherer, P. E., Williams, S., Fogliano, M., Baldini, G. & Lodish, H. F. A novel serum protein similar to C1q, produced exclusively in adipocytes. J. Biol. Chem. 270, 26746–26749 (1995)
    Article CAS Google Scholar
  6. Hu, E., Liang, P. & Spiegelman, B. M. AdipoQ is a novel adipose-specific gene dysregulated in obesity. J. Biol. Chem. 271, 10697–10703 (1996)
    Article CAS Google Scholar
  7. Maeda, K. et al. cDNA cloning and expression of a novel adipose specific collagen-like factor, apM1 (AdiPose most abundant gene transcript 1). Biochem. Biophys. Res. Commun. 221, 286–289 (1996)
    Article CAS Google Scholar
  8. Nakano, Y., Tobe, T., Choi-Miura, N. H., Mazda, T. & Tomita, M. Isolation and characterization of GBP28, a novel gelatin-binding protein purified from human plasma. J. Biochem. 120, 803–812 (1996)
    Article CAS Google Scholar
  9. Hotta, K. et al. Plasma concentrations of a novel, adipose-specific protein, adiponectin, in type 2 diabetic patients. Arterioscler. Thromb. Vasc. Biol. 20, 1595–1599 (2000)
    Article CAS Google Scholar
  10. Yamauchi, T. et al. The fat-derived hormone adiponectin reverses insulin resistance associated with both lipoatrophy and obesity. Nature Med. 7, 941–946 (2001)
    Article CAS Google Scholar
  11. Berg, A. H., Combs, T. P., Du, X., Brownlee, M. & Scherer, P. E. The adipocyte-secreted protein Acrp30 enhances hepatic insulin action. Nature Med. 7, 947–953 (2001)
    Article CAS Google Scholar
  12. Fruebis, J. et al. Proteolytic cleavage product of 30-kDa adipocyte complement-related protein increases fatty acid oxidation in muscle and causes weight loss in mice. Proc. Natl Acad. Sci. USA 98, 2005–2010 (2001)
    Article ADS CAS Google Scholar
  13. Yamauchi, T. et al. Adiponectin stimulates glucose utilization and fatty-acid oxidation by activating AMP-activated protein kinase. Nature Med. 8, 1288–1295 (2002)
    Article CAS Google Scholar
  14. Tomas, E. et al. Enhanced muscle fat oxidation and glucose transport by ACRP30 globular domain: acetyl-CoA carboxylase inhibition and AMP-activated protein kinase activation. Proc. Natl Acad. Sci. USA 99, 16309–16313 (2002)
    Article ADS CAS Google Scholar
  15. Kahn, B. B., Alquier, T., Carling, D. & Hardie, D. G. AMP-activated protein kinase: ancient energy gauge provides clues to modern understanding of metabolism. Cell Metab. 1, 15–25 (2005)
    Article CAS Google Scholar
  16. Kersten, S., Desvergne, B. & Wahli, W. Roles of PPARs in health and disease. Nature 405, 421–424 (2000)
    Article ADS CAS Google Scholar
  17. Yamauchi, T. et al. Globular adiponectin protected ob/ob mice from diabetes and apoE deficient mice from atherosclerosis. J. Biol. Chem. 278, 2461–2468 (2003)
    Article CAS Google Scholar
  18. Yamauchi, T. et al. Cloning of adiponectin receptors that mediate antidiabetic metabolic effects. Nature 423, 762–769 (2003)
    Article ADS CAS Google Scholar
  19. Wess, J. G-protein-coupled receptors: molecular mechanisms involved in receptor activation and selectivity of G-protein recognition. FASEB J. 11, 346–354 (1997)
    Article CAS Google Scholar
  20. Yamauchi, T. et al. Targeted disruption of AdipoR1 and AdipoR2 causes abrogation of adiponectin binding and metabolic actions. Nature Med. 13, 332–339 (2007)
    Article CAS Google Scholar
  21. Iwabu, M. et al. Adiponectin and AdipoR1 regulate PGC-1α and mitochondria by Ca2+ and AMPK/SIRT1. Nature 464, 1313–1319 (2010)
    Article ADS CAS Google Scholar
  22. Richter, E. A. & Ruderman, N. B. AMPK and the biochemistry of exercise: implications for human health and disease. Biochem. J. 418, 261–275 (2009)
    Article CAS Google Scholar
  23. Wu, Z. et al. Mechanisms controlling mitochondrial biogenesis and respiration through the thermogenic coactivator PGC-1. Cell 98, 115–124 (1999)
    Article CAS Google Scholar
  24. Handschin, C. & Spiegelman, B. M. The role of exercise and PGC1α in inflammation and chronic disease. Nature 454, 463–469 (2008)
    Article ADS CAS Google Scholar
  25. Cantó, C. et al. AMPK regulates energy expenditure by modulating NAD+ metabolism and SIRT1 activity. Nature 458, 1056–1060 (2009)
    Article ADS Google Scholar
  26. Paffenbarger, R. S., Jr et al. The association of changes in physical-activity level and other lifestyle characteristics with mortality among men. N. Engl. J. Med. 328, 538–545 (1993)
    Article Google Scholar
  27. Open Innovation Center for Drug Discovery http://www.ocdd.u-tokyo.ac.jp/library_e.html (The University of Tokyo, 2012)
  28. Hawley, S. A. et al. Characterization of the AMP-activated protein kinase kinase from rat liver and identification of threonine 172 as the major site at which it phosphorylates AMP-activated protein kinase. J. Biol. Chem. 271, 27879–27887 (1996)
    Article CAS Google Scholar
  29. Mootha, V. K. et al. Errα and Gabpa/b specify PGC-1α-dependent oxidative phosphorylation gene expression that is altered in diabetic muscle. Proc. Natl Acad. Sci. USA 101, 6570–6575 (2004)
    Article ADS CAS Google Scholar
  30. Berchtold, M. W. et al. Calcium ion in skeletal muscle: its crucial role for muscle function, plasticity, and disease. Physiol. Rev. 80, 1215–1265 (2000)
    Article CAS Google Scholar
  31. Shulman, G. I. Cellular mechanisms of insulin resistance. J. Clin. Invest. 106, 171–176 (2000)
    Article CAS Google Scholar
  32. Brownlee, M. Biochemistry and molecular cell biology of diabetic complications. Nature 414, 813–820 (2001)
    Article ADS CAS Google Scholar
  33. Lochhead, P. A. et al. 5-aminoimidazole-4-carboxamide riboside mimics the effects of insulin on the expression of the 2 key gluconeogenic genes PEPCK and glucose-6-phosphatase. Diabetes 49, 896–903 (2000)
    Article CAS Google Scholar
  34. Hotamisligil, G. S., Shargill, N. S. & Spiegelman, B. M. Adipose expression of tumor necrosis factor-alpha: direct role in obesity-linked insulin resistance. Science 259, 87–91 (1993)
    Article ADS CAS Google Scholar
  35. Wellen, K. E. & Hotamisligil, G. S. Inflammation, stress, and diabetes. J. Clin. Invest. 115, 1111–1119 (2005)
    Article CAS Google Scholar
  36. Weisberg, S. P. et al. Obesity is associated with macrophage accumulation in adipose tissue. J. Clin. Invest. 112, 1796–1808 (2003)
    Article CAS Google Scholar
  37. Xu, H. et al. Chronic inflammation in fat plays a crucial role in the development of obesity-related insulin resistance. J. Clin. Invest. 112, 1821–1830 (2003)
    Article CAS Google Scholar
  38. Lumeng, C. N., Bodzin, J. L. & Saltiel, A. R. Obesity induces a phenotypic switch in adipose tissue macrophage polarization. J. Clin. Invest. 117, 175–184 (2007)
    Article CAS Google Scholar
  39. Zhang, H. M. et al. Geldanamycin derivative ameliorates high fat diet-induced renal failure in diabetes. PLoS ONE 7, e32746 (2012)
    Article ADS CAS Google Scholar
  40. Li, S., Shin, H. J., Ding, E. L. & van Dam, R. M. Adiponectin levels and risk of type 2 diabetes: a systematic review and meta-analysis. J. Am. Med. Assoc. 302, 179–188 (2009)
    Article CAS Google Scholar
  41. Pischon, T. et al. Plasma adiponectin levels and risk of myocardial infarction in men. J. Am. Med. Assoc. 291, 1730–1737 (2004)
    Article CAS Google Scholar
  42. Pajvani, U. B. et al. Complex distribution, not absolute amount of adiponectin, correlates with thiazolidinedione-mediated improvement in insulin sensitivity. J. Biol. Chem. 279, 12152–12162 (2004)
    Article CAS Google Scholar
  43. Luo, Z., Saha, A. K., Xiang, X. & Ruderman, N. B. AMPK, the metabolic syndrome and cancer. Trends Pharmacol. Sci. 26, 69–76 (2005)
    Article CAS Google Scholar

Download references

Acknowledgements

We thank N. Kubota, K. Hara, I. Takamoto, Y. Hada, T. Kobori, H. Umematsu, S. Odawara, T. Aoyama, Y. Jing, S. Wei, K. Soeda and H. Waki for technical help and support; and K. Miyata, Y. Nishibaba, M. Yuasa and A. Hayashi for technical assistance and support. This work was supported by a Grant-in-aid for Scientific Research (S) (20229008, 25221307) (to T.K.), Grant-in-aid for Young Scientists (A) (23689048) (to M.I.), Targeted Proteins Research Program (to T.K.), the Global COE Research Program (to T.K.), Translational Systems Biology and Medicine Initiative (to T.K.) and Translational Research Network Program (to M.O.-I.) from the Ministry of Education, Culture, Sports, Science and Technology of Japan. Funding Program for Next Generation World-Leading Researchers (NEXT Program) (to T.Y.) from Cabinet Office, Government of Japan.

Author information

Author notes

  1. Miki Okada-Iwabu, Toshimasa Yamauchi and Masato Iwabu: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Diabetes and Metabolic Diseases, Graduate School of Medicine, The University of Tokyo, Tokyo 113-0033, Japan
    Miki Okada-Iwabu, Toshimasa Yamauchi, Masato Iwabu, Ken-ichi Hamagami, Koichi Matsuda, Mamiko Yamaguchi, Kohjiro Ueki & Takashi Kadowaki
  2. Department of Integrated Molecular Science on Metabolic Diseases, 22nd Century Medical and Research Center, The University of Tokyo, Tokyo 113-0033, Japan
    Miki Okada-Iwabu, Toshimasa Yamauchi, Masato Iwabu & Takashi Kadowaki
  3. Department of Molecular Medicinal Sciences on Metabolic Regulation, 22nd Century Medical and Research Center, The University of Tokyo, Tokyo 113-0033, Japan
    Miki Okada-Iwabu, Toshimasa Yamauchi & Takashi Kadowaki
  4. RIKEN Systems and Structural Biology Center, Tsurumi, Yokohama 230-0045, Japan,
    Teruki Honma, Hiroaki Tanabe, Tomomi Kimura-Someya, Mikako Shirouzu, Akiko Tanaka & Shigeyuki Yokoyama
  5. Graduate School of Comprehensive Human Sciences, University of Tsukuba, Tsukuba 305-8577, Japan
    Hitomi Ogata & Kumpei Tokuyama
  6. Open Innovation Center for Drug Discovery, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan,
    Tetsuo Nagano & Akiko Tanaka
  7. Graduate School of Science, The University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan,
    Shigeyuki Yokoyama

Authors

  1. Miki Okada-Iwabu
    You can also search for this author inPubMed Google Scholar
  2. Toshimasa Yamauchi
    You can also search for this author inPubMed Google Scholar
  3. Masato Iwabu
    You can also search for this author inPubMed Google Scholar
  4. Teruki Honma
    You can also search for this author inPubMed Google Scholar
  5. Ken-ichi Hamagami
    You can also search for this author inPubMed Google Scholar
  6. Koichi Matsuda
    You can also search for this author inPubMed Google Scholar
  7. Mamiko Yamaguchi
    You can also search for this author inPubMed Google Scholar
  8. Hiroaki Tanabe
    You can also search for this author inPubMed Google Scholar
  9. Tomomi Kimura-Someya
    You can also search for this author inPubMed Google Scholar
  10. Mikako Shirouzu
    You can also search for this author inPubMed Google Scholar
  11. Hitomi Ogata
    You can also search for this author inPubMed Google Scholar
  12. Kumpei Tokuyama
    You can also search for this author inPubMed Google Scholar
  13. Kohjiro Ueki
    You can also search for this author inPubMed Google Scholar
  14. Tetsuo Nagano
    You can also search for this author inPubMed Google Scholar
  15. Akiko Tanaka
    You can also search for this author inPubMed Google Scholar
  16. Shigeyuki Yokoyama
    You can also search for this author inPubMed Google Scholar
  17. Takashi Kadowaki
    You can also search for this author inPubMed Google Scholar

Contributions

M.O.-I., M.I., T.Y., T.H., K.-i.-H., K.M., M.Y., H.T., T.K-S., M.S., H.O., K.T. and A.T. performed experiments. T.K., T.Y., M.O.-I. and M.I. conceived the study. T.K., A.T., T.Y. and S.Y. supervised the study. T.Y., T.K., M.O.-I. and M.I. wrote the paper. All authors interpreted data.

Corresponding authors

Correspondence toToshimasa Yamauchi or Takashi Kadowaki.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Extended data figures and tables

Extended Data Figure 1 Phosphorylation of AMPK in C2C12 myotubes.

Phosphorylation of AMPK normalized to the amount of AMPK in C2C12 myotubes treated for 5 min with 15 µg ml−1 adiponectin or the indicated small-molecule compounds (10 μM). #, AdipoRon; ##, no. 112254; ###, no. 165073.

Extended Data Figure 2 Distribution curves showing Z scores.

a, Distribution curve showing Z scores representing AMPK activity for all compounds tested in C2C12 myotubes shown in Extended Data Table 1 and Extended Data Fig. 1. The dashed line indicates the Z score cut-off for compounds scored as hits, which showed higher activity than 80% of that seen with adiponectin. b, Distribution curve showing Z scores representing AdipoR dependency of AMPK activation for 39 compounds tested in C2C12 myotubes shown in Extended Data Table 2. Indicated are the location of AdipoRon, another hit (no. 112254), and non-hit (no. 165073).

Extended Data Figure 3 The effect of AdipoRon on complex I activity, and expression of Adipor1 and Adipor2 mRNA in C2C12 myotubes transfected with the indicated siRNA duplex.

a, Complex I activities were measured with the indicated concentrations of rotenone or AdipoRon. b, c, Adipor1 (b) and Adipor2 (c) mRNA levels were analysed by RT–qPCR. All values are presented as mean ± s.e.m. a, n = 3–7; b, c, n = 3 each; *P < 0.05 and **P < 0.01 compared to control or unrelated siRNA cells. NS, not significant.

Extended Data Figure 4 AdipoRon binding to AdipoR1 and AdipoR2.

a–d, Binding and Scatchard analyses of [3H]AdipoRon to primary hepatocytes from wild-type (a), _Adipor2_−/− knockout (b), _Adipor1_−/− knockout (c) and _Adipor1_−/− _Adipor2_−/− double-knockout (d) mice. e–h, Concentration-dependent competitive [3H]AdipoRon binding to primary hepatocytes from wild-type (e), _Adipor2_−/− knockout (f), _Adipor1_−/− knockout (g) and _Adipor1_−/− _Adipor2_−/− double-knockout (h) mice. Binding analyses were performed using the indicated concentrations of AdipoRon. c.p.m., counts per minute.

Extended Data Figure 5 Raw data of Fig. 2 and time course of glucose-lowering effect of AdipoRon.

a–m, Raw data of Fig. 2a (a), Fig. 2d, left (b, c), Fig. 2d, right (d, e), Fig. 2e, left (f, g), Fig. 2e, right (h, i), Fig. 2g, left (j, k) and Fig. 2g, right (l, m). n, Time course of glucose-lowering effect of AdipoRon. Data are calculated from data in Fig. 4a. The glucose-lowering effect of AdipoRon was obtained by the following equation and expressed as %: (vehicle plasma glucose − AdipoRon plasma glucose)/vehicle plasma glucose. All values are presented as mean ± s.e.m.

Extended Data Figure 6 The effects of compounds 112254 and 165073 on insulin resistance and glucose intolerance via AdipoR.

a, b, Chemical structures of compounds 112254 (a) and 165073 (b). c–j, Plasma glucose (c left, d left, f, g left, h left, j), plasma insulin (c right, d right, g right, h right) and insulin resistance index (e, i) during oral glucose tolerance test (OGTT) (1.0 g glucose per kg body weight) (c, d, g, h) or during insulin tolerance test (ITT) (0.5 U insulin per kg body weight) (f, j), in wild-type and _Adipor1_−/− _Adipor2_−/− double-knockout mice, treated with or without compounds 112254 or 165073 (50 mg per kg body weight). All values are presented as mean ± s.e.m. c–f, n = 10 each; g–j, n = 7 each from 2, 3 independent experiments, *P < 0.05 and **P < 0.01 compared to control or as indicated. NS, not significant.

Extended Data Figure 7 The effects of AdipoRon on glucose metabolism in _Adipor1_−/−, _Adipor2_−/− and _Adipor1_−/− _Adipor2_−/− mice.

a, Triglyceride content (a) and TBARS (b) in skeletal muscle from wild-type or _Adipor1_−/− _Adipor2_−/− double-knockout mice treated with or without AdipoRon (50 mg per kg body weight). c–g, The effects of AdipoRon on glucose metabolism in _Adipor1_−/−, _Adipor2_−/− and _Adipor1_−/− _Adipor2_−/− mice. Plasma glucose (c–f, left panels), plasma insulin (c–f, right panels) and insulin resistance index (g) during oral glucose tolerance test (OGTT) (1.0 g glucose per kg body weight). All values are presented as mean ± s.e.m. a–d, f, n = 10 each; e, n = 7 each; g, n = 7–10; *P < 0.05 and **P < 0.01 compared to vehicle mice. NS, not significant.

Extended Data Figure 8 Chemical structures and AdipoR dependency of AMPK activation.

a–d, Chemical structures of AdipoRon (a), compound 168198 (b), compound 112254 (c) and compound 103694 (d). Within the 1-benzyl 4-substituted 6-membered cyclic amine moiety, the cyclic amine moiety is surrounded by a dashed red circle, and the aromatic ring is surrounded by a light green circle. Cyan and dark green circles surround the carbonyl group and the terminal aromatic ring, respectively, located on the opposite side from the benzyl cyclic amine. e, Phosphorylation and amount of AMPK in C2C12 myotubes treated for 5 min with the indicated small-molecule compounds. Phosphorylation and amount of AMPK in C2C12 myotubes, treated for 5 min with the indicated small-molecule compounds (10 μM) (% relative to adiponectin). f, AdipoR dependency of AMPK activation. Phosphorylation and amount of AMPK in C2C12 myotubes and transfected with or without the AdipoR1 siRNA duplex, treated for 5 min with the indicated small molecule. AdipoR-dependency ratios were obtained by the following equation: 100 − (ratio for those transfected with the AdipoR1 siRNA duplex/ratio for those transfected without the AdipoR1 siRNA duplex) × 100 (%).

Extended Data Table 1 Values of phosphorylation of AMPK in C2C12 myotubes

Full size table

Extended Data Table 2 Phosphorylation of AMPK in AdipoR knock-down C2C12 myotubes

Full size table

Supplementary information

PowerPoint slides

Rights and permissions

About this article

Cite this article

Okada-Iwabu, M., Yamauchi, T., Iwabu, M. et al. A small-molecule AdipoR agonist for type 2 diabetes and short life in obesity.Nature 503, 493–499 (2013). https://doi.org/10.1038/nature12656

Download citation