TURNING PEPTIDES INTO DRUGS Background Materials & Methods 1) Bataille et al., FEBS Letters, 146, 79-86, ) Gros et al., Endocrinology, 133, , ) Wynne et al., Diabetes, 54, , Results Species Dose (nmol/kg) T ½ i.v. (hours) T ½ s.c. (hours) T max (hours) F sc (%) Mouse (N=3) Minipig (N=2) ± ± ± ± 2 ZP2929 is a selective and full agonist on hGluR and hGLP-1R ZP2929 has a pharmacokinetic profile compatible with once daily dosing in humans. Fig. 3. Effect of s.c. administration of the Glu-GLP-1 agonist ZP2929 and Exendin-4 on the change in body weight over the 6 weeks treatment period in HFD fed C57BL/6J mice. Data are mean ± SEM. ***P< compared to vehicle. (N=9-10). Fig. 2. Effect of s.c. administration of exendin-4 and the Glu-GLP-1 agonist ZP2929 on blood glucose during an OGTT (A) and Area under the Curve (AUC) (B) in HFD fed C57BL/6J mice. Data are mean ± SEM. * p<0.05. (N=10). Fig. 4. Effect of s.c. administration of the Glu-GLP-1 agonist ZP2929 on HDL/LDL ratio (A) and triglyceride concentration (B) after 6 weeks treatment in HFD fed C57BL/6J mice. Data are mean + SEM. *p<0.05, **p<0.01, *** p<0.001 compared to vehicle. (N=8-10). The new dual Glucagon-GLP-1 agonist ZP2929 improves oral glucose tolerance and reduces body weight in long-term high fat fed mice. Additionally, ZP2929 decreases body weight gain and improves HDL/LDL ratio as well as triglyceride concentration in mice fed a high fat diet for 4 weeks. Based on data from mice and monkeys, the PK profile of ZP2929 is compatible with once daily dosing in humans. The New Dual Glucagon-GLP-1 Agonist ZP2929 Improves Glycemic Control and Reduces Body Weight in Murine Models of Obesity and Type 2 Diabetes Jens R. Daugaard, Eddi Meier, Ditte Riber, Camilla Æ. Bæk, Kjeld S. Larsen and Gita Kampen Zealand Pharma A/S, Smedeland 36, DK-2600 Glostrup, Denmark Peptide hGLP- 1R (nM) hGLP-1R (SEM R ) hGluR (nM) hGluR (SEM R ) hGLP- 1R (%) hGlu R (%) GLP (1.8)> (1.0)1000 Exendin (1.1)> (1.0)1000 Liraglutide0.18(1.3)22(1.3)991 Glucagon2.3(1.2)0.027(1.1)199 Oxyntomodulin1.1(1.0)0.63(1.1)3763 ZP (1.0)0.16(1.1)5842 Aim ZP2929 significantly improves oral glucose tolerance in 30 week HFD mice ZP2929 significantly decreases body weight gain in 4 week HFD mice Results ZP2929 significantly improves HDL/LDL ratio and Triglyceride concentration in 4 week HFD mice Fig. 1. Effect of s.c. administration of exendin-4 and the Glu-GLP-1 agonist ZP2929 on body weight in HFD fed C57BL/6J mice. Data are mean ± SEM. (N=10). ZP2929 significantly decreases body weight in 30 week HFD mice ZP2929 is a dual Glucagon-GLP-1 agonist developed to provide a novel once daily treatment option for people with type 2 diabetes. ZP2929 targets glycemic control with superior and sustained weight loss as a key add-on benefit. The biological rationale for developing ZP2929 is based on the pharmacology of the gut peptide hormone oxyntomodulin. Oxyntomodulin is released from the L cells of the small intestine in response to meal ingestion, and is believed to exert its biological effects by activating both the GLP-1 receptor and the glucagon receptor, i.e. by acting as a dual Glucagon-GLP-1 agonist (1, 2). Human proof-of-concept for pharmacological use of oxyntomodulin has been demonstrated in overweight subjects (3). Oxyntomodulin, GLP-1 and glucagon are derived from pre-proglucagon. Zealand Pharma has used the glucagon sequence as a scaffold to develop a patentable dual glucagon-GLP-1 receptor agonist suitable for once daily treatment of people with type 2 diabetes. GluR and GLP-1R efficacy: HEK293 cells expressing hGluR or hGLP-1-R were incubated with increasing concentrations of test peptides, 100  M IBMX and 6 mM glucose for 15 min at 37  C. cAMP content was measured using the FlashPlate  cAMP kit. EC 50 and efficacies were estimated by computer aided curve fitting methods. Pharmacokinetics: Blood samples were collected from mice or monkeys at different time points following administration (s.c. or i.v) and the content of plasma ZP2929 was measured by LC-MS/MS. Pharmacokinetic parameters were estimated using non-compartmental analysis in WinNonLin V4.1. Pharmacodynamics: C57BL/6J male mice, 6 weeks old, were acclimatized in their new environment with free access to a high fat diet (HFD) and water. 36 weeks old, the animals were randomized into groups with similar average fasting (6 hours) blood glucose. The mice were treated twice daily s.c. for three weeks with exendin-4, ZP2929 or vehicle. Body weight was recorded daily. After 3 weeks of peptide treatment an oral glucose tolerance test (OGTT) was performed after subjecting the animals to a 6 hour fast. Following an initial blood sample animals were dosed with peptide or vehicle in the morning, and the OGTT was performed 2 hours later. The oral dose of glucose was given and the animals were returned to their home cages (t = 0). Blood glucose was measured at t=15 min, t=30 min, t=60 min, t=90 min and t=120 min. C57BL/6J male mice, 6 weeks old, were acclimatized in their new environment with free access to a high fat diet and water. 10 weeks old, the animals were randomized into groups with similar average body weight and treated twice daily s.c. with ZP2929, exendin-4 or vehicle. Throughout the study, body weights were recorded daily and used to administer the body weight-corrected doses of peptide. After treatment, blood samples were taken and blood lipids measured. To study the effect of the new dual Glucagon-GLP-1 agonist ZP2929 on oral glucose tolerance and body weight in murine models of type 2 diabetes and obesity. 46 th Annual Meeting of the European Association for the Study of Diabetes, September, 2010, Stockholm/Sweden References Conclusion SEM R :Statistical scattering of EC 50 values follows a logarithmic distribution and SEM R are thus estimated as the log ratio.