1. GLP-1/Glucagon dual agonist
Lipolytic, Energy Expenditure, and Insulinotropic Effects of HM12525A:
A Novel Long-Acting GLP-1/Glucagon Dual Agonist
P866
SY Jung1,, YJ Park1 , JK Kim1, JS Lee1, YM Lee1, YH Kim1, JH Kang1, M Trautmann2, M Hompesch2, SC Kwon1
1Hanmi Pharm. Co., Ltd., Seoul, South Korea, 2Profil Institute, Chula Vista, CA, USA
Improved Liver Function in NASH Animal Model
BACKGROUND
RESULTS
Figure 5. Changes in energy expenditure mediator expression by
HM12525A 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 Activation
Potential beneficial effects of GLP-1/Glucagon dual agonist
Body 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 day
Figure 1. HbA1c and body weight reduction in db/db mice (n=7, 4 weeks) 6h
UCP-1 mRNA
3T3-L1 differentiation
a) HbA1c
b) Body weight gain
3T3-L1 (White adipocyte)
GLP-1/Glucagon dual agonist
Liver
TG ↓
Ketogenesis ↑
FGF21 ↑
Brain
Food intake ↓
Fat Cells
Lipolysis ↑
Lipogenesis ↓
Energy
Expenditure ↑
Pancreas
Insulin secretion ↑
Stomach
Gastric 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 HM12525A
Figure 2. ipGTT in normal mice (n=7)
Immunogenic Potential
HM12525A increased the expression of UCP-1 in white adipocytes, suggesting
browning of WAT by HM12525A.
a) ipGTT
b) AUCipGTT
Figure 9. Ex vivo T cell activation of HM12525A
Lipid and Fat Mass Reduction by Lipolytic Activity
Immunogenic threshold
HM12525A is a long acting GLP-1/Glucagon dual agonist with
balanced dual agonism at GLP-1 and Glucagon receptors
Figure 6. Body composition change in normal and DIO mice (n=10, 4 weeks)
Body fat mass
Lean body mass
Body muscle mass
Body weight
Test materials
Relative activity (cAMP assay)
GLP : Glucagon
% of GLP-1
% of Glucagon
GLP-1
100
---
Glucagon
HM12525A 30 37
1 : 1
-23 %
-48.8 %
Not significant 4%
***
*** 2%
**p<0.01, ***p<0.001 vs vehicle by Anova test
Both non-conjugated active moiety and HM12525A showed negligible T cell
activation, which is below the immunogenic threshold among 50 healthy human
donors.
HM12525A improved glucose tolerance dose-dependently in normal mice.
Not significant
Figure 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 loss
b) Cumulative food intake
HM12525A showed more potent weight loss even with less food intake
inhibition compared with liraglutide.
AIMS
Figure 10. Clinical development milestone
To assess glycemic control of HM12525A
To assess body weight and fat mass changes by HM12525A
To assess liver function improvement by HM12525A
HM12525A
2014
2015
2016
2017
2018
2019
2020
2021
NDA submission
*** ; P<0.001, vs DIO vehicle
First In Human
(SAD + MAD)
SAD : Q
MAD : Q P1
METHODS
2019
No lean mass and body muscle reduction was observed by HM12525A treatment
T2DM P2 P3
-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 adipocytes
2020
Obesity P2 P3
Epinephrine
1000 nM
HM12525A
100 nM
Vehicle
CONCLUSIONS
HM12525A 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 model
Figure 4. Energy expenditure in DIO mice (n=10, 4 weeks)
a) Energy expenditure
b) Locomotor activity
HM12525A, 100 nM
Vehicle
+ Glucagon antagonist *
REFERENCES
*Glucagon antagonist : (des His1, Glu9) GCG (1-29)
1. Diabetes (2009) 58 : 2258 ~ 2266.
2. Nat. Rev Endocrinol. (2014) 10 : 24 ~ 36
3. Diabetes (2013) 62 : 1131 ~1138
HM12525A 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, 2014
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Hanmi Pharm. Co., Ltd.