Journal: Diabetes, obesity & metabolism

Article Title: GLP-1R biased cAMP agonism maintains glycemic control with reduced malaise and emesis in preclinical mammalian models

doi: 10.1111/dom.70427

Figure Lengend Snippet: cAMP accumulation and β arr recruitment to different species of GLP1R. (A) Concentration-response curves for cAMP accumulation in response to either GLP-1 (7–36), Ex-4, or Ex-Phe1 using HEK Freestyle cells transiently expressing the indicated species-specific GLP1R construct. Data represent the mean values ± SEM of N = 3 independent experiments performed with 5 replicates at each point. (B and C) Concentration-response curves for the formation of (B) GLP1R- β arr1 or (C) GLP1R- β arr2 complexes in response to either GLP-1 (7–36), Ex-4, or Ex-Phe1 using HEK Freestyle cells transiently expressing the indicated species-specific GLP1R- β arr construct pairs. Data represent the mean values ± SEM of N = 3 independent experiments performed with 4 replicates at each point.

Article Snippet: 20-week-old male mice fed with high fat high sugar (HFHS) diet for 14 weeks were surgically implanted with Alzet pump subcutaneously for continuous delivery of Ex-4 (12.6 and 126 μg/kg/day), Ex-Phe1 (12.6 and 126 μg/kg/day) or vehicle (40 mM Tris, pH 8).

Techniques: Concentration Assay, Expressing, Construct

Journal: Diabetes, obesity & metabolism

Article Title: GLP-1R biased cAMP agonism maintains glycemic control with reduced malaise and emesis in preclinical mammalian models

doi: 10.1111/dom.70427

Figure Lengend Snippet: Effects of chronic treatment with Ex-Phe1 in HFHS mice. (A) Ex-Phe1 suppressed HFHS diet intake more robustly than Ex-4 at similar doses ( n = 5/group). (B) Ex-Phe1 reduced body weight to a greater magnitude than Ex-4 at similar doses. (C) Treatment with Ex-Phe1 (126 μg/kg) induced significant fat loss measured by NMR ( n = 5/group). All data are expressed as mean ± SEM. Data in (A, B) were analysed with two-way repeated-measures ANOVA followed by Tukey’s post hoc test ( p <0.05): * Vehicle versus Ex-4 (126 μg/kg), @Vehicle versus Ex-Phe1 (12.6 μg/kg), $ Vehicle versus Ex-Phe1 (126 μg/kg), # Vehicle versus Ex-4 (12.6 μg/kg), and Ex-4 (12.6 μg/kg) versus Ex-Phe1 (126 μg/kg), ∞ Ex-Phe1 (12.6 μg/kg) versus Ex-Phe1 (126 μg/kg), ∋ Ex-4 (126 μg/kg) versus Ex-Phe1 (126 μg/kg), ⊥ Ex-4 (12.6 μg/kg) versus Ex-4 (126 μg/kg), Δ Ex-4 (12.6 μg/kg) versus Ex-Phe1 (12.6 μg/kg). Data in (C) were analysed with one-way ANOVA followed by Tukey’s post hoc test. Means with different letters are significantly different from each other ( p <0.05).

Article Snippet: 20-week-old male mice fed with high fat high sugar (HFHS) diet for 14 weeks were surgically implanted with Alzet pump subcutaneously for continuous delivery of Ex-4 (12.6 and 126 μg/kg/day), Ex-Phe1 (12.6 and 126 μg/kg/day) or vehicle (40 mM Tris, pH 8).

Techniques:

Journal: Diabetes, obesity & metabolism

Article Title: GLP-1R biased cAMP agonism maintains glycemic control with reduced malaise and emesis in preclinical mammalian models

doi: 10.1111/dom.70427

Figure Lengend Snippet: Effects of chronic treatment with Ex-Phe1 in HFD rats. (A) Ex-Phe1 chronic administration had no effect on body weight in rats maintained on HFD ( n = 7–8/group). (B) Chronic treatment with Ex-Phe1 had no effect on HFD intake in rats ( n = 7–8/group). (C) Ex-Phe1 chronic administration did not induce kaolin intake (a proxy for nausea in rodents). (D) Ex-Phe1 treatment had similar potency to Ex-4 at enhancing glycemic control during an OGTT in HFD rats ( n = 7–8/group). (E) AUC analyses of blood glucose from 0 to 120 min during an OGTT. All data expressed as mean ± SEM. (F) Chronic, continuously-delivered Ex-Phe1 reduced body weight and noncumulative food intake (G) to a lesser magnitude and duration than Ex-4 in rats maintained on HFD ( n = 5–8/group). (H) Similarly, Continuously-delivered Ex-Phe1 and Ex-4 induced kaolin intake for a shorter duration than Ex-4 after minipump implantation in rats maintained on HFD ( n = 5–8/group). (I) Acute treatment with Ex-Phe1 induces a hyperglycemic response similar to Ex-4 during an OGTT in HFD rats ( n = 12). (J) AUC analyses of blood glucose from 0 to 120 min during an OGTT. (A)–(I) Analysed with two-way repeated-measures ANOVA followed by Tukey’s post hoc test ( p <0.05): *Vehicle versus Ex-4, #Ex-Phe1 versus Ex-4, @Vehicle versus Ex-Phe1. (E), (J) Analysed with one-way repeated-measures ANOVA followed by Tukey’s post hoc test. Means with different letters are significantly different ( p <0.05).

Article Snippet: 20-week-old male mice fed with high fat high sugar (HFHS) diet for 14 weeks were surgically implanted with Alzet pump subcutaneously for continuous delivery of Ex-4 (12.6 and 126 μg/kg/day), Ex-Phe1 (12.6 and 126 μg/kg/day) or vehicle (40 mM Tris, pH 8).

Techniques: Control

Journal: Diabetes, obesity & metabolism

Article Title: GLP-1R biased cAMP agonism maintains glycemic control with reduced malaise and emesis in preclinical mammalian models

doi: 10.1111/dom.70427

Figure Lengend Snippet: Effects of acute treatment with Ex-Phe1 in musk shrews. (A) Ex-4 dose-dependently reduces body weight and (B) suppresses food intake. (C) The number of single emetic episodes in 120 min following systemic administration of vehicle and various doses of Ex-4. (D) Ex-Phe1 modestly reduces body weight and (E) suppresses food intake at 210 μg/kg and 2.1 mg/kg, however to a lesser magnitude than Ex-4. (F) The number of single emetic episodes in 120 min following systemic administration of vehicle and various doses of Ex-Phe1. (G) Ex-Phe1 and Ex-4 show equal potency in enhancing glucose clearance during IPGTT (2 g/kg). (H) AUC analyses from 0 to 120 min. Ex-4 and Ex-Phe1 treated animals had lower AUC 0–120 compared to vehicle treated animals. For (A)–(E) n = 10, for (F), (G) n = 9). All data are expressed as mean ± SEM. In the emetic studies, the number of animals exhibiting emesis, expressed as a fraction of the total number of animals tested, is indicated above each treatment group. Data in (A)–(G) were analysed with two-way repeated-measures ANOVA followed by Tukey’s post hoc test. Data in (H) were analysed with one-way repeated measures ANOVA followed by Tukey’s post hoc test. Means with different letters are significantly different from each other ( p <0.05). Vehicle versus 21 μg/kg Ex-Phe1: * p <0.05, *** p <0.0001; Vehicle versus 210 μg/kg Ex-Phe1: ### p <0.0001; Vehicle versus 21 μg/kg Ex-4: @@@ p <0.0001; Vehicle versus 210 μg/kg Ex-4: ∋ p <0.05, ∋∋∋ p <0.001; 210 μg/kg Ex-Phe1 versus 21 μg/kg Ex-4: ∞ p <0.05.

Article Snippet: 20-week-old male mice fed with high fat high sugar (HFHS) diet for 14 weeks were surgically implanted with Alzet pump subcutaneously for continuous delivery of Ex-4 (12.6 and 126 μg/kg/day), Ex-Phe1 (12.6 and 126 μg/kg/day) or vehicle (40 mM Tris, pH 8).

Techniques:

Journal: Diabetes, obesity & metabolism

Article Title: GLP-1R biased cAMP agonism maintains glycemic control with reduced malaise and emesis in preclinical mammalian models

doi: 10.1111/dom.70427

Figure Lengend Snippet: (A)–(C) Representative immunofluorescent images showing c-Fos positive cells in the area postrema (AP) and nucleus of the solitary tract (NTS) after vehicle (saline), Ex-4 (84 μg/kg, I.P.), or Ex-Phe1 (84 μg/kg, I.P.) in mice ( n = 5/group). (D) Quantification of c-Fos-positive neurons in the AP and NTS of mice. (E)–(G) Representative immunofluorescent images showing c-Fos positive cells in the AP and NTS after vehicle (saline), Ex-4 (3 μg/kg, I.P.), or Ex-Phe1 (3 μg/kg, I.P.) in rats ( n = 10/group). (I)–(K) Representative immunofluorescent images showing c-Fos positive cells in the AP and NTS after vehicle (saline), Ex-4 (210 μg/kg, I.P.), or Ex-Phe1 (210 μg/kg, I.P.) in shrews ( n = 3–4/group). Data in (D)–(L) analysed using unpaired Studenťs t -tests. All data are expressed as mean ± SEM. Means with different letters are significantly different from each other ( p <0.05). Images in (A)–(C) were acquired at 10× magnification. Images in (E)–(K) were acquired at 20× magnification. (A)–(C) Scale bar= 200 μm.

Article Snippet: 20-week-old male mice fed with high fat high sugar (HFHS) diet for 14 weeks were surgically implanted with Alzet pump subcutaneously for continuous delivery of Ex-4 (12.6 and 126 μg/kg/day), Ex-Phe1 (12.6 and 126 μg/kg/day) or vehicle (40 mM Tris, pH 8).

Techniques: Saline