<b>Table 3.</b> VO2, EE and activity during the dark cycle in calorimetry experiments.

<p>In all VO2 and EE measurements above, adjustment for activity rendered VO2 and EE either significantly higher in the TTP2515 group or equivalent between groups. W, Watts; Values are mean ± SE.</p>**<p>p<0.01,</p>*<p>p<0.05 vs water.</p

Effects of AgRP and TTP2515 on α-MSH-induced cAMP production.

<p>(<b>A)</b> α-MSH-induced cAMP production in HEK293 cells overexpressing human MC4R (green diamond); inhibition of α-MSH(3 nM)-induced cAMP production by increasing amounts of AgRP (pink diamond) (<b>B)</b> α-MSH-induced cAMP production in HEK293 cells overexpressing human MC4-R in the presence of 20 nM AgRP, 3 nM α-MSH and increasing concentrations of TTP2515 (red diamond). (C+, cAMP production in the presence of α-MSH (3 nM) and AgRP (20 nM) and in the absence of TTP2515. C-, cAMP production in the presence of α-MSH (3 nM) no AgRP or TTP2515).</p

Effects of TTP2515 on metabolic and calorimetry parameters in AgRP KO and WT mice on a 45% high fat diet.

<p>(<b>A–F</b>) AgRP KO and WT mice received TTP2515 (30 mg/kg) and AgRP KO mice received water twice daily via oral gavage; all groups were on a 45% fat diet. (<b>A</b>) During days 1–4, there was an overall suppressive effect of TTP2515 treatment on food intake (p<0.05). Food intake was lower on days 2–4 in the WT TTP2515 and KO TTP2515 groups compared to the KO water group (p<0.05) (<b>B</b>) WT TTP2515 mice gained significantly less fat mass during treatment vs. the KO water group, however, this was not true for the KO TTP2515 group (p = 0.12). (<b>C</b>) Overall, there was a significant suppressive effect of treatment on cumulative body weight change (p = 0.01); body weight gain was more frequently significantly lower in the WT TTP2515 vs. the KO water group, rather than in the KO TTP2515 vs. the KO water group (<b>D,E</b>) Total and free T4 levels were elevated in both TTP2515 treatment groups compared to the KO water group. (<b>F–H</b>) AgRP KO mice were administered either TTP2515 (30 mg/kg days 1–7; 50 mg/kg days 7–10) or water twice daily. (<b>F,G</b>) On day 3 during the dark cycle, oxygen consumption and activity were significantly lower in TTP2515–treated mice. (<b>H</b>) RQ was not different between groups during the light or dark cycle at the 50 mg/kg dose (showing day 10, representative graph of this timeperiod). +p<0.05 KO Water vs KO TTP2515; *p<0.05 KO Water vs WT TTP2515; ^#p<0.05 vs water.</p

Effects of TTP2515 in WT or AgRP KO mice acutely switched to a very high-fat diet.

<p>(<b>A–F</b>) Mice were switched from breeder chow to a 60% fat diet and started treatment with TTP2515 (30 mg/kg) or water twice daily. (<b>A–C</b>) Cumulative food intake, cumulative weight gain and fat mass were all significantly lower in TTP2515 treated mice. (<b>D</b>) Total T4 levels were significantly higher in TTP2515 treated mice. (<b>E</b>) Leptin levels tended to be lower in TTP2515 treated mice (p = 0.12). (<b>F</b>) Insulin levels were similar between groups. (<b>G–J</b>) In a separate experiment, AgRP KO mice on a 10% fat chow diet were switched to a 60% fat diet and started treatment with TTP2515 (5, 15, or 30 mg/kg) or water twice daily. (<b>G,I</b>) Cumulative caloric intake and cumulative weight gain were dose-dependently lower in TTP2515 treated mice compared to the water group. (<b>H</b>) Fat mass was lower in TTP2515 treated mice at both the 15 and 30 mg/kg doses vs. water. The 5 mg/kg group tended (p = 0.05) to have lower fat mass than the water group. (<b>J</b>) After 5 days of treatment, total T4 levels were equivalent between groups, however fasting and continued treatment revealed an increase in total T4 levels. *p<0.05, **p<0.01 vs water; <i>a</i>p<0.05 vs water, <i>b</i>p<0.05 vs 5 mg/kg, <i>c</i>p<0.05 vs 15 mg/kg.</p

<b>Table 1.</b> Effects of TTP2515 on hormone levels and liver weight in rats receiving icv AgRP infusion.

<p>All measurements reported at sacrifice in <i>ad lib</i> fed rats. Values are mean ± SE.</p>*<p>p<0.05 all groups;</p>†<p>p<0.05 vs AgRP+Water.</p

Effects of TTP2515 on refeeding and T4 after fasting in AgRP KO and WT mice.

<p>(<b>A–F</b>) AgRP KO and WT mice were fasted for 24 h and received either water or TTP2515 at 10, 25, or 50 mg/kg in three different experiments. (<b>A,B</b>) At the 10 mg/kg dose, no differences in food intake or body weight gain were observed. (<b>C,D</b>) At the 25 mg/kg dose, there was an overall significant attenuation in food intake (p<0.05) and weight gain (p<0.01) in the WT TTP2515 vs. the WT water group. Post-hoc analysis revealed that this was significant at the 20 h for food intake and at 9 and 20 h for weight gain. No differences in food intake or weight gain were observed between the KO TTP2515 vs KO water group. (<b>E</b>) At the 50 mg/kg dose, there was an overall significant attenuation in food intake in both the WT TTP2515 (p<0.01) and KO TTP2515 (p<0.05) groups vs. their water controls. Final cumulative food intake at the 20 h timepoint was significantly less in both TTP2515 groups vs. their water groups. (<b>F</b>) At the 50 mg/kg dose, final cumulative weight gain at 20 h was significantly less only in the WT TTP2515 vs WT water group. (<b>G</b>) A separate group of mice was fasted and blood samples were obtained before and after treatment. No difference in total T4 levels were observed before or after treatment. ***p<0.0001, **p<0.01, *p<0.05 vs respective water control.</p

Effects of TTP2515 in DIO and leptin-deficient mice.

<p>All measurements reported at sacrifice in <i>ad lib</i> fed mice unless otherwise indicated.</p><p>Values are mean ± SE.</p>*<p>p<0.05 vs water.</p

Effects of TTP2515 on metabolic and calorimetry parameters in leptin-deficient mice.

<p>Leptin-deficient mice received either TTP2515 at increasing doses (5–50 mg/kg) or water twice daily via oral gavage. (<b>A</b>) Food intake tended to be lower in TTP2515 treated mice during the 15 mg/kg treatment period and was significantly lower during the 30 and 50 mg/kg treatment periods. (<b>B</b>) Fat mass was significantly decreased after 24 days of TTP2515 treatment, while lean mass was not different between groups. (<b>C</b>) Body weight was significantly lower in TTP2515-treated mice on the 50 mg/kg dose. (<b>D</b>) Immediately upon starting the 50 mg/kg dose, mean respiratory quotient was significantly decreased in TTP2515 treated mice during the light cycle. (<b>E</b>) Total T4 levels at sacrifice were unchanged between groups. +p = 0.06, *p<0.05, **p<0.01 vs water.</p

Effects of TTP2515 on metabolic and calorimetry parameters in DIO mice.

<p>(<b>A–E</b>) DIO mice (16 weeks on 45% fat diet) received TTP2515 (30 mg/kg) or water twice daily via oral gavage. (<b>A,B</b>) TTP2515 treatment significantly decreased food intake and body weight gain. (<b>C</b>) Percent fat mass was significantly lower in TTP2515 treated mice at the end of the study. (<b>D</b>) TTP2515 treated mice had elevated total T4 levels. (<b>E</b>) Total T3 levels were similar between groups. (<b>F–H</b>) In a separate experiment DIO mice (45% fat diet for 15 weeks) received TTP2515 (30 mg/kg) or water twice daily via oral gavage. (<b>F,G</b>) Mean oxygen consumption and total activity were significantly decreased during the dark cycle of day 2 in TTP2515 treated mice. (<b>H</b>) Mean respiratory quotient was significantly decreased during the first half of the light cycle of day 3 in TTP2515 treated mice. **p<0.0001, *p<0.05 vs. water.</p