Presentation on theme: "GLP-1/Glucagon dual agonist"— Presentation transcript:
1GLP-1/Glucagon dual agonist Lipolytic, Energy Expenditure, and Insulinotropic Effects of HM12525A:A Novel Long-Acting GLP-1/Glucagon Dual AgonistP866SY Jung1,, YJ Park1 , JK Kim1, JS Lee1, YM Lee1, YH Kim1, JH Kang1, M Trautmann2, M Hompesch2, SC Kwon11Hanmi Pharm. Co., Ltd., Seoul, South Korea, 2Profil Institute, Chula Vista, CA, USAImproved Liver Function in NASH Animal ModelBACKGROUNDRESULTSFigure 5. Changes in energy expenditure mediator expression byHM12525A in white adipocytes (3T3-L1 cells)Figure 8. Liver function improvement in MCD-diet db/db mice (12 days)Improved Glycemic Control by GLP-1R ActivationPotential beneficial effects of GLP-1/Glucagon dual agonistBody weight(MCD + db/db mice, n=7)Liver weight(MCD + db/db mice, n=7)SREPB-1c mRNA level(MCD + db/db mice, n=3-6)Energy expenditure & Lipolysis Insulin secretion & Food intake ~9 dayFigure 1. HbA1c and body weight reduction in db/db mice (n=7, 4 weeks)6hUCP-1 mRNA3T3-L1 differentiationa) HbA1cb) Body weight gain3T3-L1 (White adipocyte)GLP-1/Glucagon dual agonistLiverTG ↓Ketogenesis ↑FGF21 ↑BrainFood intake ↓Fat CellsLipolysis ↑Lipogenesis ↓EnergyExpenditure ↑PancreasInsulin secretion ↑StomachGastric emptying ↓22.4%(+9.3 g)13.9%(+5.7 g)14.5%(+5.7 g)******UCP1 mRNA in 3T3-L1 cells (Mouse adipocyte, Q-PCR)*****1.0%(+0.4 g)**p<0.01, ***p<0.001 vs vehicle by Anova test** p<0.01, *** p<0.001 vs vehicle***HM12525A decreased HbA1c as well as body weight gain in db/db mice.HM12525A showed reduction of liver weight and SREPB-1c mRNA level in MCD-diet db/db mice which indicates NASH could be improved by HM12525AFigure 2. ipGTT in normal mice (n=7)Immunogenic PotentialHM12525A increased the expression of UCP-1 in white adipocytes, suggestingbrowning of WAT by HM12525A.a) ipGTTb) AUCipGTTFigure 9. Ex vivo T cell activation of HM12525ALipid and Fat Mass Reduction by Lipolytic ActivityImmunogenic thresholdHM12525A is a long acting GLP-1/Glucagon dual agonist withbalanced dual agonism at GLP-1 and Glucagon receptorsFigure 6. Body composition change in normal and DIO mice (n=10, 4 weeks)Body fat massLean body massBody muscle massBody weightTest materialsRelative activity (cAMP assay)GLP : Glucagon% of GLP-1% of GlucagonGLP-1100---GlucagonHM12525A30371 : 1-23 %-48.8 %Not significant4%******2%**p<0.01, ***p<0.001 vs vehicle by Anova testBoth non-conjugated active moiety and HM12525A showed negligible T cellactivation, which is below the immunogenic threshold among 50 healthy humandonors.HM12525A improved glucose tolerance dose-dependently in normal mice.Not significantFigure 3. Body weight loss in DIO mice (n=6, 2 weeks)Improved BWL by Increase of Energy Expenditure-1%-10%-17%-31%a) Body weight lossb) Cumulative food intakeHM12525A showed more potent weight loss even with less food intakeinhibition compared with liraglutide.AIMSFigure 10. Clinical development milestoneTo assess glycemic control of HM12525ATo assess body weight and fat mass changes by HM12525ATo assess liver function improvement by HM12525AHM12525A20142015201620172018201920202021NDA submission*** ; P<0.001, vs DIO vehicleFirst In Human(SAD + MAD)SAD : QMAD : QP1METHODS2019No lean mass and body muscle reduction was observed by HM12525A treatmentT2DMP2P3-36%Db/db mice (n=7) were treated (s.c) with HM12525A once a week, or liraglutide once daily, for 4 weeks respectively. Blood glucose levels were measured using a glucometer.For ipGTT, overnight fasted C57BL/6J mice were administrated with either HM12525A or liraglutide (i.v.), and the 2 g/kg of glucose was subsequently administrated (i.p.). Blood glucose and the serum insulin level was determined at 0, 30, 60, 120 min using commercially available kit.26 weeks HFD induced C57BL/6J mice (n=6) were treated (s.c) with HM12525A once a week, or with liraglutide once daily for 2 weeks respectively. The body weight and food intake was monitored daily.Energy Expenditure as well as home-cage activity were assessed by using a combined indirect calorimetry system for 1 week after the first administration in DIO mice. O2 consumption and CO2 production were measured every 10 min for a total of 7 days to determine the respiratory quotient and energy expenditure. Home-cage locomotor activity was determined using a multidimensional infrared light beam system. .To differentiate into adipocytes, confluent 3T3-L1 cells (day 1) were incubated with 0.5 mM IBMX, 0.5 μM dexamathasone, and 1 μg/ml insulin added to DMEM/10% FBS for 2 days. On day 3, the medium was changed to DMEM/FBS 10% with 1 μg/ml insulin, which was repeated every 2 days. The cells were fully differentiated into adipocytes after 10 to 14 days incubation of differentiation medium. HM12525A was co-incubated in the presence or absence of GCGR antagonist from day 3 to the end of experiment. On day 14, to evaluate the effects of HM12525A on lipid droplet formation, Oil-Red O staining was conducted. Briefly, the cells were fixed with 4% paraformaldehyde for 30 min, followed by incubation of Oil-Red O solution for 1 hr. After washing, Oil-Red O stained lipid droplet was determined using optical microscope-51%Figure 7. Reduction of lipid droplet formation in 3T3-L1 adipocytes2020ObesityP2P3Epinephrine1000 nMHM12525A100 nMVehicleCONCLUSIONSHM12525A showed glucose lowering efficacy in normal and db/db mice by balanced activity on both GLP-1 and glucagon receptors.Potent body weight loss was driven by fat mass reduction and the lean/muscle mass did not changed, because of increased energy expenditure.HM12525A showed therapeutic potential in NASH animal modelFigure 4. Energy expenditure in DIO mice (n=10, 4 weeks)a) Energy expenditureb) Locomotor activityHM12525A, 100 nMVehicle+ Glucagon antagonist *REFERENCES*Glucagon antagonist : (des His1, Glu9) GCG (1-29)1. Diabetes (2009) 58 : 2258 ~ 2266.2. Nat. Rev Endocrinol. (2014) 10 : 24 ~ 363. Diabetes (2013) 62 : 1131 ~1138HM12525A reduced lipid droplet formation in 3T3-L1 adipocytes via glucagon action.HM12525A increased energy expenditure without locomotor activity change.European Association for the Study of Diabetes 50th Annual Meeting; Vienna, Austria; September 15-19, 2014For any questions, please contact Hanmi Pharm. Co., Ltd.,Phone: ;HanmiHanmi Pharm. Co., Ltd.