Adiponectin secreted from adipocytes binds to adiponectin receptors AdipoR1 and AdipoR2, and exerts antidiabetic effects via activation of AMPK and PPAR-α pathways, respectively. Levels of adiponectin in plasma are reduced in obesity, which causes insulin resistance and type 2 diabetes. Thus, orally active small molecules that bind to and activate AdipoR1 and AdipoR2 could ameliorate obesity-related diseases such as type 2 diabetes. Here we report the identification of orally active synthetic small-molecule AdipoR agonists. One of these compounds, AdipoR agonist (AdipoRon), bound to both AdipoR1 and AdipoR2 in vitro. AdipoRon showed very similar effects to adiponectin in muscle and liver, such as activation of AMPK and PPAR-α pathways, and ameliorated insulin resistance and glucose intolerance in mice fed a high-fat diet, which was completely obliterated in AdipoR1 and AdipoR2 double-knockout mice. Moreover, AdipoRon ameliorated diabetes of genetically obese rodent model db/db mice, and prolonged the shortened lifespan of db/db mice on a high-fat diet. Thus, orally active AdipoR agonists such as AdipoRon are a promising therapeutic approach for the treatment of obesity-related diseases such as type 2 diabetes.
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Extended data figures and tables
Extended Data Figures
- Extended Data Figure 1: Phosphorylation of AMPK in C2C12 myotubes. (430 KB)
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. (51 KB)
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. (145 KB)
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. (247 KB)
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. (210 KB)
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. (251 KB)
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. (241 KB)
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. (124 KB)
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 Tables
- Supplementary Information (719 KB)
This file contains Supplementary Results, Text and Data and additional references.