Volume 23, Issue 3, Pages 427-440 (March 2016) A Long-Acting FGF21 Molecule, PF-05231023, Decreases Body Weight and Improves Lipid Profile in Non-human Primates and Type 2 Diabetic Subjects  Saswata Talukdar, Yingjiang Zhou, Dongmei Li, Michelle Rossulek, Jennifer Dong, Veena Somayaji, Yan Weng, Ronald Clark, Adhiraj Lanba, Bryn M. Owen, Martin B. Brenner, Jeffrey K. Trimmer, Kathryn E. Gropp, Jeffrey R. Chabot, Derek M. Erion, Timothy P. Rolph, Bryan Goodwin, Roberto A. Calle  Cell Metabolism  Volume 23, Issue 3, Pages 427-440 (March 2016) DOI: 10.1016/j.cmet.2016.02.001 Copyright © 2016 Elsevier Inc. Terms and Conditions

Cell Metabolism 2016 23, 427-440DOI: (10.1016/j.cmet.2016.02.001) Copyright © 2016 Elsevier Inc. Terms and Conditions

Figure 1 PF-05231023 Decreases Body Weight and Food Intake in Obese NHPs (A) NHP study design. Male obese drug-naive cynomolgus monkeys weighing between 7–16 kg were included in this study for the indicated periods. The first dose of PF-05231023 was administered on day 1. Animals were randomized based on BW, and six animals were included in each dose group. (B and C) Changes from the baseline upon treatment with PF-05231023 (n = 6/group) for (B) BW and (C) food intake. The mean body weight for all groups on day 1 was 10.7 ± 0.95 kg, and the mean daily food intake in week 1 prior to dosing for all groups was 236 ± 9.8 g. (D) Mean food intake in the morning prior to dosing (9 a.m.) and in the evening at 6 p.m. in animals treated with vehicle (n = 5) and 10 mg/kg PF-05231023 (n = 18). (E) Percent change of food intake from days 1–7 in animals treated with vehicle (n = 5) and 10 mg/kg PF-05231023 (n = 18). Food intake data do not include the consumption of a daily apple. (F) BW change from the baseline in pair-fed animals and animals administered 10 mg/kg PF-05231023 (n = 18) for vehicle (n = 6) and pair-fed (n = 6). ∗p < 0.05 using one-way ANOVA and Dunnett’s post hoc test. Statistical analyses for BW and food intake are provided in Table S1. Cell Metabolism 2016 23, 427-440DOI: (10.1016/j.cmet.2016.02.001) Copyright © 2016 Elsevier Inc. Terms and Conditions

Figure 2 PF-05231023 Decreases Adiposity and Improves the Lipid Profile in Obese NHPs (A) Mean difference of abdominal circumference (centimeters) on day 29 compared with the baseline of the indicated dose groups. Veh, vehicle. (B) Percent change of axial adiposity by DEXA scan on day 29 compared with the baseline. (C) Percent change in bone mineral content (BMC) using a DEXA scan. (D) Representative histology images from adipose tissue in animals administered vehicle or the indicated doses of PF-05231023. (E) LSMean ± SEM percent change of fasting TGs. (F) Fasting BHBA. (G) Percent change of circulating adiponectin on day 29 compared with the baseline in animals treated with vehicle (n = 6) and 10 mg/kg PF05231023 (n = 6). (H and I) Percent change from the baseline in leptin on hay 8 (H) and day 29 (I) in animals treated with vehicle (n = 5) and 10 mg/kg PF-05231023 (n = 6). NS, not significant. (J) Plasma lipoprotein profiles in animals treated with vehicle and 10 mg/kg PF-05231023 on the indicated days. n = 6 animals per group (A–C) for all dose groups except 10 mg/kg (n = 18). ∗p < 0.05. Statistical analyses for TGs are reported in Table S1. Cell Metabolism 2016 23, 427-440DOI: (10.1016/j.cmet.2016.02.001) Copyright © 2016 Elsevier Inc. Terms and Conditions

Figure 3 PF-05231023 Modulates Gene Expression in Subcutaneous Adipose Tissue of NHPs SC adipose tissue was collected on day −10 (baseline) and day 29 from NHPs dosed with vehicle (n = 6) or PF-05231023 (n = 18). (A–N) FGFR1 (A), KLB (B), CD36 (C), Adiponectin (D), AdipoR (E), PPARγ (F), leptin (G), LepRb (H), Dio2 (I), IL10 (J), IL1β (K), IFNγ (L), CCL2 or MCP-1 (M), and fatty acid binding protein 4 (aP2) (N). Relative mRNA for the indicated genes was normalized to the expression of TBP1. ∗p < 0.05, #p < 0.09, determined by paired t test. Cell Metabolism 2016 23, 427-440DOI: (10.1016/j.cmet.2016.02.001) Copyright © 2016 Elsevier Inc. Terms and Conditions

Figure 4 The effect of PF-05231023 in Type 2 Diabetic Subjects (A) Phase 1b clinical study schematic. (B–G) Percent change from baseline of (B) BW, (C) fasting TGs, (D) TC, (E) LDL, (F) HDL, and (G) adiponectin. Significance is indicated as follows: ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 on day 25 compared with placebo. All data are LSMeans ± SEM. Statistical analyses are reported in Table S2. Cell Metabolism 2016 23, 427-440DOI: (10.1016/j.cmet.2016.02.001) Copyright © 2016 Elsevier Inc. Terms and Conditions

Figure 5 Pharmacodynamic Effects of PF-05231023 in Type 2 Diabetic Subjects (A–H) Percent change from the baseline in mean daily glucose (MDG) (A), fasting plasma insulin (milliunits per liter) (B), free (C) and total (D) IGF-1 (nanograms per milliliter), CTX (nanograms per milliliter) (E), osteocalcin (nanograms per milliliter) (F), P1NP (nanograms per milliliter) (G), and BSAP (nanograms per milliliter) (H). ∗p < 0.05, ∗∗p < 0.01 on day 25 compared with placebo. Statistical analyses are reported in Table S2. Cell Metabolism 2016 23, 427-440DOI: (10.1016/j.cmet.2016.02.001) Copyright © 2016 Elsevier Inc. Terms and Conditions