0 7 14 21 28 35 42 49 56 631
10
100
0.1
1
10
100
LAPSCA-Exendin-4 3mg
LAPSInsulin 115 1.5nmol/kg
LA
PSC
A-Exen
din
-4 s
eru
m c
on
c. (n
M)
Time (day)
LA
PS In
au
lin
115 s
eru
m c
on
c. (n
M)
Switching from daily basal insulin to weekly LAPSInsulin 115 + LAPSCA-Exendin-4 combination showed improved efficacy and body weight gain reduction.
0 1 24
6
8
10
12
14Vehicle
Insulin glargine 18.5nmol/kg
Insulin glargine 18.5nmol/kg -> LAPSInsulin 115 4.4nmol/kg
Insulin glargine 18.5nmol/kg (BID) -> HM12470 4.4nmol/kg + HM11260C 0.72nmol/kg (Q2D)
Hb
A1c (
%)
4
6
8
10
12
14Vehicle
HM12470 4.4nmol/kg + HM11260C 0.36nmol/kg (Q2D)
Insulin glargine 18.5nmol/kg (BID)
Insulin glargine 18.5nmol/kg (BID) -> HM12470 4.4nmol/kg (Q2D)
Insulin glargine 18.5nmol/kg -> LAPSInsulin 115 4.4nmol/kg + LAPSCA-Exendin-4 0.72nmol/kg
Hb
A1c (%
)
(A) Time course changes in HbA1c
0 1 2 3 4 5 6 7
-3
-2
-1
0
H
bA1c
(%)
vs.
vehi
cle
Body weight (g)
LAPSCombo(LAPSInsulin 115 + LAPSCA-Exendin-4)
IGlar
0
IGlar (350U/wk as HED)
IGlar
2 4 6 wk
db/db mice
Experimental Design
LAPSInsulin 115 + LAPSCA-Exendin-4(280U/wk + 1mg/wk as HED)
IGlar LAPS Insulin 115(280U/wk as HED)
Switch
0 7 14 21 28 35 42-3
-2
-1
0
1
H
bA
1c (
%vs. V
eh
icle
)
Time (day)
-0.8 LAPSInsulin 115
0 7 14 21 28 35 42-3
-2
-1
0
1
H
bA1c
(%
vs.
Veh
icle
)
Time (day)
IGlar -2.5 LAPSCombo ( LAPSInsulin 115 + LAPSCA-Exendin-4)
***
***
† Mean difference vs. Vehicle at EOT
Vehicle
0 2 4 6 80.1
1
10
100
LAPSInsulin 115 only
Combination formulation
Time (Days)
LA
PS In
su
lin
115 S
eru
m C
on
c.
(nM
)
0 3 6 9 12 15 18 21
0.1
1
10
100
0.1
1
10
100
LA
PSC
A-E
xd
4 S
eru
m C
on
c. (n
M)L
AP
S Insu
lin
115 S
eru
m C
on
c. (n
M)
Time (days)
0 7 14 21 28 35 42 49 56 630.1
1
10
0.1
1
10
LAPSCA-Exendin-4 3mg
LA
PS
CA-E
xen
din
-4 seru
m co
nc. (n
M)
Time (day)
LA
PS
In
au
lin
115 seru
m co
nc. (n
M)
t1/2 = 153hrs, PTR = 1.4
t1/2 = 132hrs, PTR = 1.6
0 7 14 21 28 35 42 49 56 630.1
1
10
0.1
1
10
LAPSInsulin 115 1.5nmol/kg
LA
PS
CA
-E
xendin-4 serum
conc. (nM
)
Time (day)
LA
PS
Inaulin 115 serum
conc. (nM
)
Predicted PK in human
0 2 4 6 80.1
1
10
100
LAPSCA-Exdendin-4 only
Combination formulation
Time (Days)
LA
PS C
A-E
xd
-4 S
eru
m C
on
c. (n
M)
5
6
7
8
9
10
11
12Vehicle
LAPS-Exd4 0.36
LAPS-INS A15 8.8
LAPS-Exd4 + LAPS-INS A15 2.2
HbA1
c (%)
***
10.5
8.0
9.0
7.2
††0 1 2 3 4 5 6 7 8 9 10
0.01
0.1
1
10
100
1
10
LAPSInsulin 115 5.5 nmol/kgLAPSCA-Exendin-4 3.75 nmol/kg
Time (days)
LA
PS
-IN
S A
15
C
on
c. (n
M)
LA
PS
-E
xd
-4 C
on
c. (n
M)
† PTR : 1.4
† PTR : 1.6
†Peak-to-Through Ratio
For any questions, please contact Hanmi Pharm. Co., Ltd., Phone: +82-31-371-5141; [email protected]
Pharmacological Evaluation of Once-weekly Combination Of A Long-acting Insulin Analog With A Long-acting Exendin-4 Analog In An Animal Model
SC Kwon1, SY Jung1 , CK Lim1, YJ Park1, JK Kim1, IY Choi1, SH Lee1, YH Kim1, JH Kang1, M Trautmann2, M Hompesch2
1Hanmi Pharm. Co., Ltd., Seoul, South Korea, 2Profil Institute, Chula Vista, CA, USA
P972
RESULTS
CONCLUSIONS LAPSInsulin 115 and LAPSCA-Exendin-4 showed well-harmonized and
prolonged PK profiles compared to daily Insulin and GLP-1RA.
In a co-formulation, LAPSInsulin 115 and LAPSCA-Exendin-4 showed no PK profiles and intrinsic activity interferences.
LAPSInsulin 115 + LAPSCA-Exendin-4 combination showed superior glycemic control and reduced body weight gain in db/db mice and DIO/STZ rats.
Switching from daily basal insulin to weekly LAPSInsulin 115 + LAPSCA-Exendin-4 combination demonstrated improved glycemic and body weight control in db/db mice.
Beneficial Effects of LAPSInsulin 115 + LAPSCA-Exendin-4 Combination
Figure 1. PK comparison of weekly LAPS products vs. daily comparator in normal rats (n=3, s.c.)
Weekly Potential of LAPSInsulin 115 + LAPSCA-Exendin-4 Combination
0 2 4 6 80.1
1
10
100
LAPSInsulin 115 5.5nmol/kg
Co-formulation
Time (Days)
LA
PS
In
su
lin
115 S
eru
m C
on
c. (n
M)
0 2 4 6 8 100.01
0.1
1
10
LAPSCA-Exendin-4 3.75nmol/kg
Co-formulation
Time (Days)
LA
PS
CA
-E
xd
-4 S
eru
m C
on
c. (n
M)
Figure 3. PK profile of co-formulation vs. individual drug in normal rats (n=6, s.c., single)
LAPSInsulin 115 and LAPSCA-Exendin-4 did not show drug-drug interaction when co-formulated.
(B)
Figure 4. HbA1c (A) and Body weight changes (B) by combination treatment in db/db mice (n=5, s.c., Q2D, 5 weeks)
European Association for the Study of Diabetes 50th Annual Meeting; Vienna, Austria; September 15-19, 2014
Figure 2. Multiple dose PK of co-formulated LAPSInsulin 115 and LAPSCA-Exendin-4 in normal rats (n=5, s.c., Q3D)
Similar and prolonged PK profiles of LAPSInsulin 115 and LAPSCA-Exendin-4 were observed in SD rat and weekly human PK simulation.
0
5
10
15Vehicle
LAPS-Exd4
LAPS-INS A15 8.8
LAPS-Exd4 + LAPS-INS A15 2.2
B
ody w
eight
(g)
6.4
12.1
4.8
7.0
***
†
BWGNeutralization
LAPSInsulin 115 + LAPSCA-Exendin-4 combination showed superior efficacy with BWG neutralization by insulin dose reduction.
Figure 7. Development plan of QUANTUM project
QUANTUM is a proprietary name of Hanmi’s diabetes & obesity pipeline
No PK and Pharmacologic Drug-Drug Interaction Between LAPSInsulin 115 and LAPSCA-Exendin-4 When Co-formulated
(A) HbA1c at EOT (B) BW changes at EOT
Hanmi Hanmi Pharm. Co., Ltd.
REFERENCES1. Clinical Therapeutics (2013) 35: 714–232. Nature Review Endocrinology (2012) 8: 728-42
3. Diabetologia (2014) 50 (Suppl. 1):P9334. PLoS ONE (2010) 5: e9540
***p<0.001 vs. vehicle by ANOVA test†p<0.05, † † p<0.01 by ANOVA test
Table 1. in vitro pharmacologic activity of co-formulation
The co-formulation showed no interference with the intrinsic activity of individual drugs
Formulation% Activity vs. LAPSInsulin 115 % Activity vs. LAPSCA-Exendin-4
Insulin-Rbinding
Insulin-R phos-phorylation
GLP-1R binding
cAMP accumula-tion
LAPSInsulin 115 100% 100% - -
LAPSCA-Exendin-4 - - 100% 100%
Co-formulation (1:0.3*)
117% 105% 106.7% 95.9%
Co-formulation (1:8.1*)
98.8% 129% 101.2% 104.4%
Drug Interference No No No No
* Molar ratio between LAPSCA-Exendin-4 and LAPSInsulin 115 in co-formulation
Figure 5. HbA1c (A) and body weight changes (B) daily vs. weekly combination in DIO/STZ rats (n=5, s.c., BID or Q3D, 4 weeks)
0 2 4 6 80.1
1
10
100
0.1
1
10
LAPSInsulin 115 5.5 nmol/kg (weekly)LAPSCA-Exd-4 2.5 nmol/kg (weekly)
IDegludec 55.8 nmol/kg (daily)
Liraglutide 50 nmol/kg (daily)
Time (days)L
AP
S Insu
lin
115 S
eru
m C
on
c. (n
M)
LA
PSCA-E
xd
-4 S
eru
m C
on
c. (n
M)
0 2 4 6 80.1
1
10
100
0.1
1
10
LAPSInsulin 115 5.5 nmol/kg (weekly)LAPSCA-Exendin-4 2.5 nmol/kg (weekly)
IDegludec 55.8 nmol/kg (daily)
Liraglutide 50.0 nmol/kg (daily)
Time (days)
LA
PS
In
su
lin
115 S
eru
m C
on
c. (n
M)
LA
PS
CA-E
xd
-4 S
eru
m C
on
c. (n
M)
4 fold reduced insulin dose
BACKGROUND
AIMS
METHODSIn vitro pharmacology of single formulation
Receptor binding affinity of LAPSInsulin 115 and LAPSCA-Exendin-4 in co-formulation was deter-mined by SPA (scintillation proximity assay) and SPR (surface plasmon resonance) analysis, respectively. Phosphorylation of insulin receptor was determined using commercially available kit after 10min treatment of either rh-Insulin or LAPSInsulin 115 in hIR-B/CHO cells. cAMP was assayed after 10 min treatment with LAPSCA-Exendin-4 in hGLP-1R/CHO cells. All results were normalized with LAPSInsulin 115 or LAPSCA-Exendin-4.
Pharmacokinetic prediction in humanHuman serum concentration vs time of LAPSInsulin 115 was predicted by Css-MRT method us-ing PK parameters from mice, rats and dogs. Human CL was derived from rule of exponent methods, and human Vd was from allometry applied correction factor. Single-dosing PK pa-rameter from P1 study of LAPSCA-Exendin-4 were utilized for the human PK simulation.
PK analysis of LAPSInsulin 115 and + LAPSCA-Exendin-4 combinationSerum concentration of test articles were determined using the a modified ELISA, and PK pa-rameters were calculated by a non-compartmental method. In multiple-dosing PK study, LAPSIn-sulin 115 and/or LAPSCA-Exendin-4 were administrated on a Q3D interval to mimic human QW dosing.
Efficacy study in db/db mice or DIO/STZ ratsLAPSInsulin 115 and/or LAPSCA-Exendin-4 were administrated to db/db mice or DIO/STZ rats with Q2D or Q3D dosing. HbA1c level was measured after 4~6 wk treatment. Body weight change was monitored every 2 days.
To evaluate the once-weekly potential of LAPSInsulin 115 + LAPSCA-Exendin-4 combination.
Statistical analysisThe HbA1c, body weight changes by LAPSInsulin 115, LAPSCA-Exendin-4, and their combination were analyzed by one way ANOVA with Dunnett’s post test.
Beneficial effects of daily basal insulin and GLP-1RA combination
Superior glycemic control Improved safety profile (BW gain ↓, Hypoglycemic risk ↓)
Basal Insulin
Strong FBG controlAchieve A1c target 50~60%
Body weight gainHypoglycemic risk ↑
GLP-1RA
Modest FBG or PPG controlAchieve A1c target 40~60%
Body weight lossNo Hypoglycemic risk
Complementary action and insulin dose reduction
Synergistic action
Efficacy
Safety
*Image was modified from Clin Ther. 35, 714-23 (2013) and Nat Rev Endocrinol 8, 728-42 (2012)
Weekly basal insulin and GLP-1RA combination enabled by LAPS technology
LAPSCA-Exendin-4 [Ph2b, US & EU] Super-Agonist [Potent A1c Reduction & BWL] Flexible regimen from weekly to monthly Low Immunogenicity
LAPSInsulin 115 [EASD Poster #933] Low peak-to-trough ratio results in constant insulin levels
To evaluate the beneficial effects of LAPSInsulin 115 + LAPSCA-Exendin-4 combination in in vivo disease models.
LAPSInsulin 115 and LAPSCA-Exendin-4 showed similar and prolonged PK profiles compared with daily insulin and GLP-1RA, respectively.
(A)
Figure 6. ΔHbA1c (A), and relationship between ΔHbA1c and ΔBW (B) after switching from daily insulin to weekly combination in db/db mice (n=7, s.c., BID or Q2D)
Switching from Daily Insulin to a Weekly Combination Improved Ef-ficacy and Safety
0
30
60
90
120
B
od
y W
eig
ht
(g
)
71
100
38
6254
†
(A) HbA1c at EOT (B) BW changes at EOT
LAPSInsulin 115 + LAPSCA-Exendin-4 combination showed superior efficacy compared with daily combination without additional BWG.
***p<0.001 vs. vehicle by ANOVA test†p<0.05, † † p<0.01 by ANOVA test
*
3
4
5
6
7H
bA1c
(%)
***
6.6
5.4
6.3
4.8
5.4
††
**
(B) Relationship between ΔHbA1c and ΔBW
5
6
7
8
9
10
11
12
Vehicle
LAPSCA-Exendin-4 0.36 nmol/kg + LAPSInsulin 115 2.2 nmol/kg
LAPSCA-Exd4 0.36 nmol/kg + LAPSInsulin 115 8.8 nmol/kg
Legend
Hb
A1c (%
)
5
6
7
8
9
10
11
12
VehicleLAPSInsulin 115 4.04 nmol/kg ( 50U/day * 7 in human)
LAPSCA-Exendin-4 0.36 nmol/kg + LAPSInsulin 115 2.2 nmol/kg
LAPSCA-Exd4 0.36 nmol/kg + LAPSInsulin 115 8.8 nmol/kg
Legend
Hb
A1
c (%
)
5
6
7
8
9
10
11
12
Vehicle
LAPSCA-Exendin-4 1.63 nmol/kg
LAPSInsulin 115 8.8 nmol/kg
LAPSInsulin 115 4.04 nmol/kg + LAPSCA-Exendin-4 1.63 nmol/kg
IDegludec 9.3 nmol/kg + Liraglutide 15 nmol/kg ( 50U/day + 1.8mg in human)
Hb
A1
c (%
)
5
6
7
8
9
10
11
12
Vehicle
LAPSCA-Exendin-4 1.63 nmol/kg
LAPSInsulin 115 8.8 nmol/kg
LAPSInsulin 115 4.04 nmol/kg + LAPSCA-Exendin-4 1.63 nmol/kg
Hb
A1
c (%
)
LAPSInsulin 115
Switch (Day 14)
5
6
7
8
9
10
11
12
Vehicle
LAPSCA-Exendin-4 0.36 nmol/kg + LAPSInsulin 115 2.2 nmol/kg
LAPSCA-Exd4 0.36 nmol/kg + LAPSInsulin 115 8.8 nmol/kg
Legend
Legend
Hb
A1
c (%
)
5
6
7
8
9
10
11
12
VehicleLAPSInsulin 115 8.8 nmol/kg
LAPSInsulin 115 2.2 nmol/kg + LAPSCA-Exendin-4 0.36 nmol/kg
Hb
A1
c (%
)
5
6
7
8
9
10
11
12
Vehicle
LAPSCA-Exendin-4 0.36 nmol/kg
LAPSInsulin 115 8.8 nmol/kg
Hb
A1c (%
)
5
6
7
8
9
10
11
12
LAPSInsulin 115 8.8 nmol/kg
LAPSCA-Exendin-4 0.36 nmol/kg + LAPSInsulin 115 2.2 nmol/kg
LAPSCA-Exd4 0.36 nmol/kg + LAPSInsulin 115 8.8 nmol/kg
LegendVehicle
Hb
A1c (%
) -0.7 IGlar
Ag
lycosylated
Fc
Ag
lycosylated
Fc
Ag
lycosylated
Fc
Ag
lycosylated
Fc
