Poster presentation at ADA in 2014

A Novel Very Long-Acting Insulin Analog (HM12470) with Potential for Once-Weekly Dosing,
Has a Favorable PK, PD and Mitogenic Proﬁle
Serum concentration of LAPSInsulin 115 and/or LAPSCA-Exendin-4 were determined
using modified ELISA. In a chronic study, LAPSInsulin 115 and/or LAPSCA-Exendin-4
were administrated to db/db mice with Q2D interval. HbA1c level was measured after
5 wk-treatment. Body weight change was monitored every 2 days.
IR-A
IR-B
IGF-1R
100
100
100
IGF-1R/IR
1
IGF-1
0.9
0.6
17,984
23,515
Glargine
42
73
579
10
AspB10
308
248
449
1.6
Insulin 115
93
86
84
0.9
IRS-1/2
t1/2 = 25.4 h
RAS/RAF
AKT
ERK1/2
1. Lower peak-to-trough ratio due to long duration
2. Patients’ adherence improvement by onceweekly administration
3. Ideal combination partner with weekly GLP-1
agonist
15
10
5
0
Control IGF-1 Human IGlar
Insulin
Asp Insulin
B10 115
Time (days)
10
5
Control IGF-1 Human IGlar
Insulin
Asp
B10
Insulin
115
8
ERK1/2
phosphorylation
100
reduced dose
0
5
10
Time (days)
-40
LAPSInsulin
in
LAPS
4
Insulin
Insulin
Insulin 115
43.1 nmol/kg
258.3 nmol/kg
43.1 nmol/kg
Insulin 115
129.2 nmol/kg
LAPS
LAPS
2
LAPS
Control IGF-1 Human IGlar
Insulin
Asp
B10
Insulin
115
 Insulin 115 triggered levels of mitogenic signals comparable with human insulin,
whereas positive controls (IGF-1 and AspB10) had significantly higher mitogenic signals.
American Diabetes Association’s (ADA) 74th Scientiﬁc Sessions, San Francisco, CA, USA; June 13-17, 2014
1/4 reduced dose
-1.0
21
35
28
Time (days)
42
56
49
4.0
***
-1.8
-3.4
-3.3
6
*p<0.05, **p<0.01 vs vehicle by Anova test
LAPS
Insulin
21.5 nmol/kg, Q2D
LAPS
Insulin
43.1 nmol/kg, Q2D
LAPS
Insulin
86.1 nmol/kg, Q2D
LAPS
Insulin 115
Insulin 115
5.5 nmol/kg, Q2D
11.1 nmol/kg, Q2D
Insulin 115
22.2 nmol/kg, Q2D
LAPS
LAPS
IDegludec
Liraglutide
LAPS
Insulin 115
LAPS
CA-Exd-4
55.8 nmol/kg (daily)
50.0 nmol/kg (daily)
5.5 nmol/kg (weekly)
2.5 nmol/kg (weekly)
10
10
1
1
0
2
4
Time (days)
 LAPSInsulin 115 showed prolonged glucose lowering effect compared to LAPSInsulin in
db/db mice at the same dose and at a even as well as 1/6 dose level (Fig 4a).
 LAPSInsulin 115 achieved a comparable HbA1c reduction only with 1/4 the dose of
LAPSInsulin (Fig 4b).
100
10
1
LAPS
Insulin 115 only
Combination formulation
0
 LAPSInsulin
8
LAPS
CA-Exd4 0.36 nmol/kg
18
†
16
14
***
12
Neutralized
12.1
10
BWG
9.2
8
6
7.0
6.4
4
4.8
2
0
*** p<0.001 Oneway ANOVA with Dunnett’s post test vs. Vehicle
††
9.0
8.0
7
***
7.2
***
6.8
1/4 reduced dose
 LAPSInsulin 115 and
weight.
† p<0.05, †† p<0.01
LAPSCA-Exendin-4
Oneway ANOVA with Dunnett’s post test
showed synergistic effects on HbA1c and body
6
0.1
8
2
4
Time (Days)
115 or
6
8
LAPSCA-Exendin-4
For any questions, please contact Hanmi Pharm. Co., Ltd., Rep. of Korea
Phone: +82-31-371-5141; [email protected]
Figure 9. Development plan of QUANTUM project
Project
LAPS
CA-Exendin-4
LAPS
Insulin115
LAPS
Insulin Combo
2013
2014
P1
P2
2015
2016
2017
2018
2019
P1
2021
NDA Submission
2020
P3
P2
P1
2020
2018
P3
P2
P3
2021
 QUANTUM is a proprietary name of Hanmi’s diabetes & obesity pipeline
CONCLUSIONS
 In vitro binding affinity on IR and IGF-1R showed that the
mitogenic potency of Insulin 115 and LAPSInsulin 115 were not
increased compared to that of human-insulin.
 Extended PK profiles and prolonged glucose lowering
efficacy of LAPSInsulin 115 were demonstrated.
LAPSInsulin 115 can
achieve a basal insulin profille suitable for once weekly use.
 The combination of LAPSInsulin 115 and LAPSCA-Exendin-4
showed synergistic effects on glycemic control with
suppressed body weight gain.
 Human PK prediction suggests that
 Easy to combine into single formulation
 No pharmacokinetic interactions
0.1
Insulin 115
(b) Body weight change
10.5
9
5
Single platform-derived combination
***
8.8 nm ol/kg +
Insulin 115
(a) HbA1c
6
 LAPSInsulin 115 could be ideal partner to LAPSCA-Exendin-4 for once-weekly
insulin/GLP-1RA combination with well harmonized PK profiles.
***
-2.6
2.2 nm ol/kg + LAPSCA-Exd4 0.36 nm ol/kg
LAPS
115 + LAPSCA-Exendin-4
100
0.1
10
63
Figure 7. PK profile of combination vs mono in single-shot
formulation (SD rats, n=6)
*
**
-2.0
LAPS
0.36 nm ol/kg
CA-Exd4
Body weight (g)
Serum conc.(nM)
8
6
Vehicle
Vehicle
6
0
15
14
Figure 6. PK profile comparison with long-acting GLP-1R
agonist in SD rats (n=3~6, s.c.)
12
8
7
Once weekly Insulin + GLP-1 combination
(b) 4-week repeated dose
1/6
200
Actin
AKT
phosphorylation
0
4
2
10
300
ERK1/2
Cell proliferation
IGF-1R
phosphorylation
0
400
AKT
Erk1/2 phosphorylation
(Fold of control)
115 as a very long-acting insulin
IGF-1R phosphorylation
(Fold of control)
Overview of
Apoptosis
Akt phosphorylation
(Fold of control)
These images are adapted from Diabetic Medicine (2013) 30:
1293–1297 and Diabetes (2011) 60 (Suppl. 1):LB11 (37-LB)
Insulin
t1/2 = 23 hrs
500
p-ERK1/2
LAPSInsulin
-20
LAPS
600
p-AKT
PI3K
0
(a) Single dose
IGF-1R
SHC
LAPS Insulin 115
t1/2 = 76.6 hrs
0.1
Peak-to-Trough Ratio =
0
LAPSInsulin
20
Figure 4. Glucose lowering efficacy in db/db mice (n=7,s.c)
p-IGF-1R
IGF-1R
3.5 nmol/kg
1
FBG (m g/dL)
Incidence of
nocturnal
hypoglycemia
Insulin
3.5 nmol/kg
FBG_ LAPS Insulin 115
8.8 nmol/kg
LAPS
11
 Pharmacokinetics in human was predicted based on PK of three different animals.
LAPSInsulin 115 was expected extended half-life and lower peak-to-trough ratio than
LAPSInsulin.
FBG_ LAPS Insulin
LAPS
Insulin 115
1.6
LAPSInsulin, t = 55 hrs
1/2
and daily insulins.
FBG_v ehicle
10
115, t1/2 = 132 hrs
Peak-to-Trough Ratio =
1
0.1
 LAPSInsulin 115 had extended half-life and duration of action than
normal pigs.
IGF-1 receptor signaling
IGF-1
3.5 nmol/kg
0.01
 Insulin 115 had slightly reduced affinities on IR-A, -B, and IGF-1R, compared with
human insulin and no preferential binding on IGF-1R or IR-A.
Day-to-day
variability
3.5 nmol/kg
PK_LAPS Insulin 115
Vehicle
LAPSInsulin
CA-Exd-4 Serum Conc. (nM)
(Degludec vs Glargine)
PK_LAPS Insulin
3.5 nmol/kg
10
Insulin 115 Serum Conc. (nM)
Relative ratio (% vs. insulin)
Figure 1. Mitogenic signaling in MCF-7 cells
PK profile comparison
115 had extended half-life than
Insulin 115
12
8
LAPSInsulin
LAPS
LAPS
Longer is safer than shorter.
Time (days)
6
FBG (mg/dL)
Table 1. Receptor binding affinities in triplicates
Prolonged PK profile
seems to be a key.
“Paradoxical Safety”
4
100
115 (HM12470)
Insulin
2
3.5 nmol/kg
100
LAPS
Once weekly basal insulin
Test materials
0
Figure 3. PK/PD in pigs (n=3, s.c.)
RESULTS
LAPSInsulin
20K PEG Insulin LAPSInsulin
t1/2 = 18.5 hrs
t1/2 = 6.7 hrs
IDeg
t1/2 = 2.9 hrs
Insulin
CA-Exd-4 Serum Conc. (nM)
 LAPSInsulin
65.1nmol/kg (we e kly)
LAPS
LAPS
 PK and PD of LAPSInsulin 115 + LAPSCA-Exendin-4 for QW combination
65.1nmol/kg (we e kly)
1000
Insulin 115 Serum Conc. (nM)
0.1
Insulin
Insulin 115
115
t1/2 = 44.1 hrs
1
Human serum concentration-time was predicted by Css-MRT method from PK
parameters of mice, rats and dogs. Human CL was derived from rule of exponent
methods, and human Vd was from simple allometry applied correction factor.
55.8 nmol/kg (daily)
LAPS
LAPSInsulin
10
 Pharmacokinetic prediction in human
65.1 nmol/kg (daily)
IDe glude c
LAPS
Serum concentration of test articles were determined using the a modified ELISA and
PK parameters were calculated by a non-compartmental method. In an acute study,
4-hr fasting blood glucose level was measured every day after s.c administration of
test articles in db/db mice. In a chronic study, HbA1c level was measured after 4-wk
administration in db/db mice with Q2D interval for human mimetic condition.
Key requirements for once weekly insulin
t1/2 = 12.5 h
115 for once weekly basal insulin
Figure 8. HbA1c and body weight change with combination
treatment in db/db mice (n=5-6, s.c., Q2D, 5 weeks)
Figure 5. Predicted pharmacokinetics in human
20K PEG Insulin
LAPS
100
HbA1c (%)
 PK and PD analysis of
LAPSInsulin
Serum conc. (nM)
Binding affinity of test materials was measured in competition between unlabeled and
125I-labeled materials on the IR- or IGF-1R/CHO membrane. Mitogenic signaling was
detected in MCF-7 cells and the phosphorylation level was analyzed by Western blot.
BACKGROUND
 Long half-life
 Flat profile
- Low variability (intra/inter patients)
 Low risk of hypoglycemic episodes
 Flexible timing/dosing
 Little or no weight gain
 CV safety
1000
 In vitro receptor binding affinity and mitogenic signaling in MCF-7
Serum conc. (nM)
The long-acting basal insulin, LAPSInsulin 115 (HM12470) has been
developed for once-weekly administration to provide for a better basal
insulin treatment option. The objective of this study was to investigate the
in vitro properties, PK/PD of LAPSInsulin 115 in normal and diabetic animal
models to evaluate once-weekly dosing potential, and combination with
once-weekly GLP-1R agonist. Insulin 115 displayed slightly reduced
binding affinity for IR-A, -B, and IGF-1R compared with human insulin. In
addition, Insulin 115 triggered mitogenic signals in MCF-7, at a level
comparable with mitogenic signals triggered by human insulin. In a PK
study, LAPSInsulin 115 when administered subcutaneously exhibited a half
life of ~ 44 hrs in normal rats, and the extended half-life was also confirme d
in other species such as mice, dogs, pigs and monkeys. The improved PK
profile contributed to a more prolonged glucose lowering efficacy in db/db
mice compared with the native human insulin conjugate ( LAPSInsulin).
Based on the results from three different species, the half-life of LAPSInsulin
115 in humans is expected to be 132 hrs and the peak-to-trough ratio was
calculated to be 1.6 for QW dosing by the Wajima Css-MRT method. This
QW potential of LAPSInsulin 115 is expected to harmonize very well with the
PK profile of the long-acting GLP-1R agonist (LAPSCA-Exendin-4) showing
potential for a well matched QW combination therapy. LAPSInsulin 115
combined with LAPSCA-Exendin-4 showed synergistic effect on HbA1c
reduction and neutralized body weight in db/db mice. These observations
suggest that LAPSInsulin 115 has a once-weekly dosing potential with a
sufficiently extended half-life and a low mitogenic risk, as well as its
promise for once-weekly insulin/GLP-1R agonist combination therapy.
HbA 1c (% )
SY Hwang1, IY Choi1, , JY Kim1, SY Jung1, DJ Kim1, YM Lee1, YH Kim1, M Trautmann2, M Hompesch2, JW Son1, SC Kwon1
1Hanmi Pharm. Co., Ltd., Seoul, South Korea, 2Proﬁl Institute, Chula Vista, CA, USA
Figure 2. PK in SD rats (n=3, s.c.)
METHODS
ABSTRACT
89-LB
10
1
0.1
0.01
LAPS
CA-Exe ndin-4 only
Combination formulation
0
2
4
Time (Days)
6
8
did not have any interference in single formulation.
REFERENCES
1.
2.
3.
4.
Diabetic Medicine (2013) 30: 1293–1297.
Diabetes (2011) 60 (Suppl. 1):LB11 (37-LB)
PLoS ONE (2010) 5: e9540.
PLoS ONE (2012) 7: e34274.
Hanmi
Hanmi Pharm. Co., Ltd.

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