Therapeutic effect of a novel long-acting GLP-1/GIP/Glucagon … · 2018-10-17 · Therapeutic effect of a novel long-acting GLP-1/GIP/Glucagon triple agonist (HM15211) in NASH and

Therapeutic effect of a novel long-acting GLP-1/GIP/Glucagon triple agonist (HM15211) in NASH and fibrosis animal models

Jung Kuk Kim, Jong Suk Lee, Eunjin Park, Dae Jin Kim, Young Hoon Kim,

and In Young Choi

Hanmi Pharm. Co., Ltd., Seoul, Republic of Korea

Presenter Disclosure

European Association for the Study of Diabetes (EASD) 54th Annual Meeting, Berlin, Germany; 1-5 Oct., 2018

Employee of Hanmi Pharm. Co., Ltd.


NASH progression

• Inflammatory tissue injury

• HSC activation

• Fibrogenic component ↑

Scar replaces damaged liver cells

Liver function failure

Fibrosis

• Obesity;

Dyslipidemia;

Insulin resistance and T2DM

Liver fat accumulation

Hepatocyte lipotoxicity

Normal NAFL NASH Cirrhosis

• Lipid peroxidation ↑

• Oxidative stress ↑

Inflammation

Inflammatory cytokine

Resident / infiltrated macrophage activation

Liver cell apoptosis


NASH progression and potential drug candidates

Drug candidates

(selected)

GLP-1RALiraglutide; semaglutide (P2, Novo)

ACC inhibitorGS-0976 (P2, Gilead)

PPAR agonistElafibranor (P2, GENFIT)

Normal NAFL NASH Cirrhosis

Note. √ Indicated compounds were used as active comparators in efficacy studies

√

ASK1 inhibitorSelonsirtib (P3, Gilead)√

FXR agonistObeticholic acid (P3, Intercept.)

GS-9674 (P2, Gilead)

√

• Lipid peroxidation ↑

• Oxidative stress ↑

Inflammation

• Inflammatory tissue injury

• HSC activation

• Fibrogenic component ↑

Scar replaces damaged liver cells

Liver function failure

Fibrosis

• Obesity;

Dyslipidemia;

Insulin resistance and T2DM

Liver fat accumulation

Hepatocyte lipotoxicity

Inflammatory cytokine

Resident / infiltrated macrophage activation

Liver cell apoptosis


LAPSCOVERY : Long Acting Peptide/Protein DiSCOVERY Technology

GLP-1/GIP/GCG

triple co-agonist

Flexible PEG Linker

Aglycosylated Fc

fragment

What is long-acting GLP-1/GIP/Glucagon triple agonist?

[General profile]

• Extended half-life (t1/2 = 42.7 ~ 55 hrs in mice; 82.8 ~ 85.7 hrs in rats)

• High glucagon (GCG) activity suitable for obesity treatment

• Balanced GLP-1 and GIP activity to neutralize hyperglycemic risk of

high GCG

• Anti-inflammatory effect by GIP activity

• Recently completed FIH clinical study in healthy obese subjects

Hanmi’s GLP-1/GIP/GCG triple co-agonist is conjugated

with a human IgG Fc fragment via flexible linker

Weight change in pair-fed controlled DIO mice

0 2 4 6 8 1 0 1 2 1 4 1 6 1 8 2 0 2 2 2 4 2 6 2 8

-4 0

-3 0

-2 0

-1 0

0

1 0

T im e (D a y s )

Bo

dy

we

igh

t c

ha

ng

e

(%

vs

.d

ay

0)

P a i r - f e d f o r l i r a g lu t id e

P a i r e - f e d f o r H M 1 5 2 1 1

V e h ic l e

L ir a g lu t id e 5 0 n m o l/k g , B ID (3 m g /d a y in h u m a n )

H M 1 5 2 1 1 1 .4 4 n m o l/k g , Q 2 D (2 m g /w k in h u m a n )

** ~ ***p<0.01 ~ 0.001 vs. vehicle by One-way ANOVA , †††p<0.001 vs. pair-fed by One-way ANOVA

0 2 4 6 8 1 0 1 2 1 4 1 6 1 8 2 0 2 2 2 4 2 6 2 8

-4 0

-3 0

-2 0

-1 0

0

1 0

T im e (D a y s )

Bo

dy

we

igh

t c

ha

ng

e

(%

vs

.d

ay

0)

-5.0 %

-18.6 %

-35.0 %

†††

-5.8 % -21.8 %

FI independent

body weight loss

n.s.


Vehicle Liraglutide HM15211

UCP-1

PGC-1α

100μm

White adipose tissue browning

0 2 4 6 8 1 0 1 2 1 4 1 6 1 8 2 0 2 2 2 4

8

1 0

1 2

1 4

1 6

En

erg

y e

xp

en

dit

ure

(k

al/

kg

/hr)

T im e (h rs )

am 08:00 pm 08:00

Enhanced energy expenditure

H+

H+ Heat ↑

White adipocyte

Beige adipocyte

Efficient weight loss by HM15211 and related MoA

***

**

Vehicle

Liraglutide 50 nmol/kg, BID (3 mg/day in human)

HM15211 1.44 nmol/kg, Q2D (2 mg/week in human)

0 2 4 6 8 1 0 1 2 1 4 1 6 1 8 2 0 2 2 2 4 2 6 2 8

-4 0

-3 0

-2 0

-1 0

0

1 0

T im e (D a y s )

Bo

dy

we

igh

t c

ha

ng

e

(%

vs

.d

ay

0)

P a i r - f e d f o r l i r a g lu t id e

P a i r e - f e d f o r H M 1 5 2 1 1

V e h ic l e



Pair-fed for liraglutide

Pair-fed for HM15211

3) Enhanced hepatic fat reduction in DIO mice

0

5 0

1 0 0

1 5 0

2 0 0

2 5 0

He

pa

tic

TG

(m

g/g

)

***

***

Vehicle Liraglutide HM15211

H&

E

100μm

Oil-r

ed

O

100μm

1) Liver preferential distribution

4 hr 48 hr 168 hr

0

1 0 0 0

2 0 0 0

3 0 0 0

HM

15

21

1 C

on

c.

(ng

/mL

fo

r S

eru

m,

ng

/g f

or T

iss

ue

)Efficient hepatic fat reduction by HM15211 and related MoA

2) Hepatic lipid metabolism reprogramming

SREBP-1C SCD1ACC1 ACC2 FAS0 .0

0 .5

1 .0

1 .5

Fo

ld i

nc

re

as

e

*

** **

CPT1 HADHA HADHBLCAD ACADVL ACOX EHHADH ACAA1PGC-1α0

2

4

6

8

Fo

ld i

nc

re

as

e

In mitochondria In peroxisome

***

*

*

*** ***

*

*

0

1 0 0 0

2 0 0 0

3 0 0 0

HM

15

21

1 C

on

c.

(ng

/mL

fo

r S

eru

m,

ng

/g f

or

Tis

su

e)

S e ru m

L iv e r

H e a rt

L u n g

L a rg e I .

S p le e n

P a n c re a s

A d ip o s e t is s u e

S m a ll I.

S to m a c h

M u s c le

0

5 0

1 0 0

1 5 0

He

pa

tic

TG

(m

g/g

)

* * *

V e h ic l e




β-oxidationDe novo lipogenesis

HM15211

Glu

cag

on Browning of WAT

: Energy expenditure ↑

Liver targeting

: Lipolysis ↑ & lipogenesis ↓

Hypothesis


Weekly triple agonist

Appetite ↓

Inflammation ↓

GL

P-1

NASH improvement: Steatosis ↓, Inflammation ↓

Insufficient for fibrosis improvement

• Body weight ↓

• Fat mass, blood lipid ↓

• Liver fat ↓

• Expected for once-weekly regimen

• Completed for P1 SAD study in healthy

obese subjects

HM15211 [Ph1, US]

Hypothesis



Glu

cag

on

Inflammation ↓

• Liver inflammation ↓

Browning of WAT


Liver targeting


GIP

Appetite ↓

Inflammation ↓

GL

P-1

NASH improvement: Steatosis ↓, inflammation↓, ballooning↓

Fibrosis improvement

• Body weight ↓


• Liver fat ↓



obese subjects

HM15211 [Ph1, US]

Hypothesis



Glu

cag

on

• Liver inflammation ↓

Browning of WAT


Liver targeting


GIP

GL

P-1

NASH improvement: Steatosis ↓, inflammation↓ ballooning↓

Fibrosis improvement

• Body weight ↓


• Liver fat ↓

BG increasing risk ↑

Appetite ↓

Inflammation ↓

INS secretion ↑

Inflammation ↓

INS secretion ↑

Hyperglycemic risk of glucagon use ↓



obese subjects

HM15211 [Ph1, US]

Objective & study strategies

HM15211, long-acting GLP-1/GIP/Glucagon triple agonist, might have therapeuticpotential in NASH and fibrosis as well as obesity


• The efficacy was evaluated in rodent disease models

C57BL/6 mice(8 weeks old) MCD-diet Lean;

NASH w/ mild to advanced fibrosis6 ~ 12 weeks

AMLN-diet28 weeks Obese; Fatty liver

Induction periods Disease status

Experimental scheme


Change of weight and steatosis score in AMLN-diet mice

0

1

2

3

NF

AL

D a

cti

vit

y s

co

re

Normal, Vehicle

AMLN-mice, Vehicle

0

1

2

3

NF

AL

D a

cti

vit

y s

co

re

Selonsertib 30 mg/kg, QD (250 mg/day in human)

Obeticholic acid 30 mg/kg, QD (250 mg/day in human)


*~***p<0.05 ~ 0.001 vs. AMLN mice, vehicle by One-way ANOVA †††p<0.001 vs.selonsirtib or OCA One-way ANOVA

Weight change (AMLN mice, n=7)

0 2 4 6 8 1 0 1 2 1 4 1 6 1 8 2 0 2 2 2 4 2 6 2 8

-4 0

-3 0

-2 0

-1 0

0

T im e (D a y s )

Bo

dy

we

igh

t c

ha

ng

e

(%

vs

.D

0)

C57BL/6 mice(8 weeks old)

Drug treatmentModel induction Analysis

28 weeksBody weight

Hepatic TG

Steatosis score4 weeks

***

†††

AMLN-diet

Experimental scheme


Change of weight and steatosis score in AMLN-diet mice

0

1

2

3

NF

AL

D a

cti

vit

y s

co

re

Normal, Vehicle

AMLN-mice, Vehicle

Steatosis score (AMLN mice, n=7)Weight change (AMLN mice, n=7)

0 2 4 6 8 1 0 1 2 1 4 1 6 1 8 2 0 2 2 2 4 2 6 2 8

-4 0

-3 0

-2 0

-1 0

0

T im e (D a y s )

Bo

dy

we

igh

t c

ha

ng

e

(%

vs

.D

0)

H&E staining (AMLN mice, representative image)

Normal, Veh AMLN, Veh Selonsertib

Obeticholic acid HM15211


Drug treatmentModel induction

AMLN-diet

Analysis

28 weeksBody weight

Hepatic TG

Steatosis score4 weeks

***

*~***p<0.05 ~ 0.001 vs. AMLN mice, vehicle by One-way ANOVA †††p<0.001 vs.selonsirtib or OCA One-way ANOVA

0

1

2

3

4

Ste

ato

sis

sc

ore

******†††

†††

0

1

2

3

NF

AL

D a

cti

vit

y s

co

re




Change of hepatic fat content in MCD-diet mice

Hepatic TG (MCD mice, n=7)

*~**p<0.05 ~ 0.01 vs. MCD mice, vehicle by One-way ANOVA; †p<0.05 vs. Liraglutide by One-way ANOVA

0

2 0

4 0

6 0

8 0

1 0 0

1 2 0

1 4 0

He

pa

tic

TG

(m

g/g

liv

er)

*

**

†


Real-time liver MRI (MCD mice, representative image)

Normal, baseline MCD, Veh, baseline MCD, Veh, week 2 HM15211 week 2

MCD, Veh, week 4 HM15211 week 4

Lira week 2

Lira week 4

Experimental scheme


Hepatic TG, TBARS

Blood liver function marker

Marker expression (qPCR, IHC)

NAS


MCD-diet

MRI MRI MRI

4 weeks6 weeks

Analysis

0

5

1 0

1 5

2 0

2 5

3 0

He

pa

tic

TB

AR

S1

) (n

mo

l/m

g)

***

*

N o r m a l m ic e , V e h ic le

M C D m ic e , V e h ic le

L ir a g lu t id e 5 0 n m o l/k g , B ID ( 3 m g /d a y in h u m a n )

H M 1 5 2 1 1 0 .3 6 n m o l/k g , Q 2 D (0 .5 m g /w k in h u m a n )


0

5

1 0

1 5

2 0

2 5

3 0

He

pa

tic

TB

AR

S1

) (n

mo

l/m

g)

***

*






0

5

1 0

1 5

2 0

2 5

3 0

He

pa

tic

TB

AR

S1

) (n

mo

l/m

g)

***

*






Normal, Vehicle

MCD mice, Vehicle


HM15211 0.72 nmol/kg, Q2D (1 mg/wk in human)


*~***p<0.05 ~ 0.001 vs. MCD mice, vehicle by One-way ANOVA ††~†††p<0.01 ~ 0.001 vs. Liraglutide by One-way ANOVA

0

2 0 0

4 0 0

6 0 0

8 0 0

1 0 0 0

1 2 0 0

Blo

od

AL

T

(IU

/L)

******

0 .0

0 .5

1 .0

1 .5

Blo

od

to

tal

bil

iru

bin

(m

g/d

L)

***

***

†††

0

5

1 0

1 5

2 0

2 5

3 0

He

pa

tic

TB

AR

S1

) (n

mo

l/m

g l

ive

r)

***

*

††

Change of NASH prognosis markers in MCD-diet mice

1) TBARS is surrogate of malondialdehyde, the lipid peroxidation product; oxidative stress marker

Hepatic TBARS1)

(MCD mice, n=7)

Blood ALT and bilirubin level (MCD mice, n=7)

0

5

1 0

1 5

2 0

2 5

3 0

He

pa

tic

TB

AR

S1

) (n

mo

l/m

g)

***

*






0

5

1 0

1 5

2 0

2 5

3 0

He

pa

tic

TB

AR

S1

) (n

mo

l/m

g)

***

*






0

5

1 0

1 5

2 0

2 5

3 0

He

pa

tic

TB

AR

S1

) (n

mo

l/m

g)

***

*






Normal, Vehicle

MCD mice, Vehicle




Change of hepatic marker expression in MCD-diet mice

Normal, vehicle MCD, vehicle

100μm

MCD, Liraglutide MCD, HM15211

F4/80 staining (MCD mice, representative image)

0

1

2

3

4

5

Re

lati

ve

ex

pre

ss

ion

(Fo

ld i

nc

re

as

e)

TNF-α

Inflammation & HSC activation marker gene expression (MCD mice, n=7, qPCR)

0

5

1 0

1 5

2 0

2 5

3 0

He

pa

tic

TB

AR

S1

) (n

mo

l/m

g)

***

*






0

5

1 0

1 5

2 0

2 5

3 0

He

pa

tic

TB

AR

S1

) (n

mo

l/m

g)

***

*






0

5

1 0

1 5

2 0

2 5

3 0

He

pa

tic

TB

AR

S1

) (n

mo

l/m

g)

***

*






Normal, Vehicle

MCD mice, Vehicle



*~***p<0.05 ~ 0.001 vs. MCD mice, vehicle by One-way ANOVA


Change of hepatic marker expression in MCD-diet mice

Normal, vehicle MCD, vehicle

100μm

MCD, Liraglutide MCD, HM15211

F4/80 staining (MCD mice, representative image)

0

1

2

3

4

5

Re

lati

ve

ex

pre

ss

ion

(Fo

ld i

nc

re

as

e)

* *** ***

TNF-α TGF-β α-SMA

Inflammation & HSC activation marker gene expression (MCD mice, n=7, qPCR)

0

5

1 0

1 5

2 0

2 5

3 0

He

pa

tic

TB

AR

S1

) (n

mo

l/m

g)

***

*






0

5

1 0

1 5

2 0

2 5

3 0

He

pa

tic

TB

AR

S1

) (n

mo

l/m

g)

***

*






0

5

1 0

1 5

2 0

2 5

3 0

He

pa

tic

TB

AR

S1

) (n

mo

l/m

g)

***

*






Normal, Vehicle

MCD mice, Vehicle





NAFLD activity score (MCD mice, n=7)

Change of NAFLD activity score in MCD-diet mice

*~**p<0.05 ~ 0.01 vs. MCD mice, vehicle by One-way ANOVA, ††p<0.01 vs. Liraglutide by One-way ANOVA

0

1

2

3

4

NA

FL

D a

cti

vit

y s

co

re

Experimental schemeModel induction

MCD-diet


0

1

2

3

4

B a llo o n in g

L o b u la r in f la m m a t io n

S t e a t o s is

NA

FL

D a

cti

vit

y s

co

re

N o rm a l

V e h c le

M C D

V e h ic le

M C D

L ira .

M C D

211

Steatosis

Lobular inflammation

Ballooning

0

5

1 0

1 5

2 0

2 5

3 0

He

pa

tic

TB

AR

S1

) (n

mo

l/m

g)

***

*






0

5

1 0

1 5

2 0

2 5

3 0H

ep

ati

c T

BA

RS

1) (

nm

ol/

mg

)

***

*






0

5

1 0

1 5

2 0

2 5

3 0

He

pa

tic

TB

AR

S1

) (n

mo

l/m

g)

***

*






Normal, Vehicle

MCD mice, Vehicle



Drug treatment

6 weeks 4 weeksLiver fat ↑

Inflammation onsetStudy #1

Expected disease status

Study #1

Veh GLP-1 Triple

**

* †††




0

5

1 0

1 5

2 0

2 5

3 0

He

pa

tic

TB

AR

S1

) (n

mo

l/m

g)

***

*






0

5

1 0

1 5

2 0

2 5

3 0H

ep

ati

c T

BA

RS

1) (

nm

ol/

mg

)

***

*






0

5

1 0

1 5

2 0

2 5

3 0

He

pa

tic

TB

AR

S1

) (n

mo

l/m

g)

***

*






Normal, Vehicle

MCD mice, Vehicle




0

1

2

3

4

NA

FL

D a

cti

vit

y s

co

re

Experimental schemeDrug treatmentModel induction

6 weeks 4 weeksMCD-diet


0

1

2

3

NF

AL

D a

cti

vit

y s

co

re



Veh GLP-1 Triple

0

1

2

3

4

B a llo o n in g


S t e a t o s is

NA

FL

D a

cti

vit

y s

co

re

N o rm a l

V e h c le

M C D

V e h ic le

M C D

L ira .

M C D

211

Steatosis


Ballooning

Study #1

10 weeks 5 weeksInflammatory liver damage ↑

liver fat ↓, ballooning ↑

Liver fat ↑


Study #2


**

* †††




0

5

1 0

1 5

2 0

2 5

3 0

He

pa

tic

TB

AR

S1

) (n

mo

l/m

g)

***

*






0

5

1 0

1 5

2 0

2 5

3 0H

ep

ati

c T

BA

RS

1) (

nm

ol/

mg

)

***

*






0

5

1 0

1 5

2 0

2 5

3 0

He

pa

tic

TB

AR

S1

) (n

mo

l/m

g)

***

*






Normal, Vehicle

MCD mice, Vehicle




0

1

2

3

4

NA

FL

D a

cti

vit

y s

co

re

Experimental schemeDrug treatmentModel induction

6 weeks 4 weeksMCD-diet


0

1

2

3

NA

FL

D a

cti

vit

y s

co

re

0.00 0.00

0

1

2

3

NF

AL

D a

cti

vit

y s

co

re



Veh ASK1i FXR TripleVeh GLP-1 Triple

0

1

2

3

4

B a llo o n in g


S t e a t o s is

NA

FL

D a

cti

vit

y s

co

re

N o rm a l

V e h c le

M C D

V e h ic le

M C D

L ira .

M C D

211

Steatosis


Ballooning

Study #1 Study #2

10 weeks 5 weeksInflammatory liver damage ↑

liver fat ↓, ballooning ↑

Liver fat ↑


Study #2


**

* †††

*** ***

*

Experimental scheme


12 weeks 4 weeks



Study #1 Study #2 Study #3


MCD-diet

Change of hepatic collagen and fibrosis score in MCD-diet mice

Hepatic hydroxyproline & fibrosis score (MCD mice, n=7)

6 weeks 4 weeks

10 weeks 5 weeks

Sirius red staining (MCD mice, representative image from study #1)

100μm

No

rma

l, v

eh

icle

MC

D, v

eh

icle

MC

D, H

M15211

0

2 0 0

4 0 0

6 0 0

8 0 0

1 0 0 0

1 2 0 0

1 4 0 0

Hy

dro

xy

pro

lin

e (

nm

ol/

g l

ive

r)

***

**

******

*

0.0 2.4 1.80.3 1.9 1.0 0.0 3.0 2.4Fibrosis

score:

0

5

1 0

1 5

2 0

2 5

3 0

He

pa

tic

TB

AR

S1

) (n

mo

l/m

g)

***

*






0

5

1 0

1 5

2 0

2 5

3 0

He

pa

tic

TB

AR

S1

) (n

mo

l/m

g)

***

*





H M 1 5 2 1 1 0 .7 2 n m o l/k g , Q 2 D (1 .0 m g /w k in h u m a n )Normal, Vehicle

MCD mice, Vehicle


Marker expression (qPCR)

Hydroxyproline

Sirius red staining

Analysis

Study #1

Study #2

Study #3

Experimental scheme


Change of hepatic fibrosis marker expression in MCD-diet mice

0

2

4

6

8

1 0

1 2

1 4

Ra

lati

ve

ex

pre

ss

ion

(Fo

ld i

nc

re

as

e)

** ** ** ***

0

2

4

1 0

1 5

2 0

Ra

lati

ve

ex

pre

ss

ion

(Fo

ld i

nc

re

as

e)

** ** ***

**

*

*~***p<0.05 ~ 0.001 vs. MCD mice, vehicle by One-way ANOVA 1) TIMP-1: Tissue Inhibitor of MetalloProtease-1

0

5

1 0

1 5

2 0

2 5

3 0

He

pa

tic

TB

AR

S1

) (n

mo

l/m

g)

***

*






0

5

1 0

1 5

2 0

2 5

3 0

He

pa

tic

TB

AR

S1

) (n

mo

l/m

g)

***

*





H M 1 5 2 1 1 0 .7 2 n m o l/k g , Q 2 D (1 .0 m g /w k in h u m a n )Normal, Vehicle

MCD mice, Vehicle


Hepatic TIMP-11) expression (MCD mice, n=7, qPCR)

Hepatic collagen-1α1 expression (MCD mice, n=7, qPCR)

Study #1 Study #2 Study #3 Study #1 Study #2 Study #3



MCD-diet

Marker expression (qPCR)

Hydroxyproline

Sirius red staining

Analysis

12 weeks 4 weeks

Study #1

Study #2

Study #3

6 weeks 4 weeks

10 weeks 5 weeks

• Considering the progression of NAFLD from simple steatosis to NASH and fibrosis, recent drug

candidates may have limited efficacy because they mainly target one step of disease progression

• In addition to efficient weight loss (energy expenditure ↑), the long-acting GLP-1/GIP/Glucagon triple agonist,

HM15211, directly reduced liver fat (lipid metabolism reprogramming) and possibly inflammation, suggestive of

therapeutic potential in NASH and fibrosis

• In AMLN-diet mice, HM15211, but not an ASK1 inhibitor and FXR agonist, provided efficient weight loss

and completely reversed steatosis

• In MCD-diet mice, HM15211 reduced both 1) liver fat, 2) oxidative stress, and 3) marker gene expression

including HSC activation (TGF-β and α-SMA) , resulting in greater NAS reduction than GLP-1RA, ASK1 inhibitor,

or a FXR agonist

• HM15211 could improve hepatic fibrosis regardless of induction period

By directly affecting key steps (lipotoxicity and inflammation), HM15211 might provide improvedtherapeutic efficacy for the treatment of NASH and fibrosis; A Clinical study in NASHpatients is planned for human efficacy translation

Summary & Conclusion

Please note posters or oral presentation reporting more information about HM15211:165-OR: Neuroprotective effects of HM15211, a novel long-acting GLP-1/GIP/Glucagon triple agonist in the neurodegenerative disease models

500-P: Bone protective effect of a novel long-acting GLP-1/GIP/Glucagon triple agonist (HM15211) in an animal model

719-P: A novel combination of a long-acting GLP-1/GIP/Glucagon triple agonist and once weekly basal insulin offers improved glucose lowering and weight loss in diabetic animal model


Neuroprotective effects of HM15211, a novel long-acting GLP-1/GIP/Glucagon triple agonist in the neurodegenerative disease models

Jeong A Kim, Sang Don Lee, Sang-Hyun Lee, Sung Min Bae, In Young Choi, and

Young Hoon Kim

Hanmi Pharm. Co., Ltd., Seoul, Republic of Korea

Employee of Hanmi Pharm. Co., Ltd.

Presenter Disclosure

European Association for the Study of Diabetes (EASD) 54th Annual Meeting, Berlin, German; 01-05 Oct., 2018

LAPSCOVERY : Long Acting Peptide/Protein DiSCOVERY Technology

GLP-1/GIP/GCG

triple agonist

Flexible PEG Linker

Aglycosylated Fc

fragment

Hanmi’s GLP-1/GIP/GCG triple agonist is conjugated with a

human IgG Fc fragment via flexible linker

[General profile]

• Extended half-life (t1/2 = 42.7 ~ 55 hrs in mice; 82.8 ~ 85.7 hrs in rats)

• High glucagon (GCG) activity suitable for obesity treatment

• Balanced GLP-1 and GIP to neutralize hyperglycemic risk of high GCG

• Anti-inflammatory effect by GIP activity

• Recently completed for FIH clinical study in healthy obese subjects

HM15211, a novel long-acting GLP-1/GIP/Glucagon triple agonist


• Neuroprotective effects of GLP-1, glucagon and GIP

• Obesity is one of the risk factors for neurological disorders

Alzheimer’s disease

↑ BMI, T2DM ↑ AD risk

Leptin/insulin resistance ↑ AD

Leptin ↓ Aβ, p-tau

Parkinson’s disease

Insulin resistance, T2DM ↑ PD

↑ Insulin levels ↑ α-synuclein aggregation

Leptin ↑ survival of DA cells

Multiple sclerosis

Obesity ↑ MS risk

Caloric restriction ↑ EAE lifespan

↓ insulin sensitivity in MS

Incretin hormones in central nervous system

GLP-1

Neuroprotection

Peripheral

contributions

Peripheral-CNS Crosstalk

↑ Neurite outgrowth

↑ Progenitor proliferation

↓ inflammation

↓ Glutamate neurotoxicity

↑ Progenitor proliferation

↓ inflammation

Glucagon

GIP


Objective

Evaluation of neuroprotective potential of HM15211…

• To assess the efficacy and related mode of actions

a. in Parkinson’s disease mice model

b. of Alzheimer’s disease in diabetic mice model


Efficacy and related MoAs in Parkinson’s disease mice model


• MPTP is a specific neurotoxin affecting the nigrostriatal system.

Glial cellBBB

Blood

MPP+

Complex I

inhibition

Dopamine

transporter

Reactive oxygen

species

Oxidative stressATP decrease

Inflammation

Cell death

6wD0

MPTP/P (twice a week)

HM15211(QW)Behavior tests

(Traction test, pole test, rotarod test)

Sacrifice

D-2

Training

for

behavior

test

C57Bl/6 ♂10 wks old

5w


Parkinson’s disease mouse model

• Experimental scheme

D7D0

MPTP

HM15211

MPTP MPTP MPTP MPTP MPTP MPTP

Behavior tests

(Traction test, pole test, rotarod test)

Sacrifice

D-1

Training

for

behavior

test

C57Bl/6

Subchronic PD model Chronic PD model

Dopaminergic neuroprotection by HM15211

Tyrosine hydroxylase (TH) :

rate limiting step for dopamine synthesis

0

5 0

1 0 0

1 5 0

TH

+ c

ell

s i

n S

ub

sta

ntia

nig

ra

V e h ic le

M P T P 3 0 m p k , ip

M P T P 3 0 m p k , ip +

L A P S -H M T 2 1 1 5 .0 3 n m o l/k g , s c

M P T P 3 0 m p k , ip +

L A P S -H M T 2 1 1 2 .5 n m o l/k g , s c

* * ** * *

* *

TH

+ a

rea

in

su

bsta

nti

a n

igra

TH

+ a

rea

in

str

iatu

m

(% v

s.

ve

hic

le)

0

5 0

1 0 0

1 5 0

TH

+ a

re

a i

n s

tria

tu

m (

% v

s.

ve

hic

le)

V e h ic le


M P T P 3 0 m p k , ip +


M P T P 3 0 m p k , ip +


* * *

* * *

*

Subchronic PD model

Chronic PD model

0

5 0

1 0 0

1 5 0

2 0 0

TH

+ c

ell

s i

n S

ub

sta

ntia

nig

ra

V e h ic le

M P T P 2 5 m g /k g (s c , tw ic e w e e k ly ) +

P ro b e n e c id 2 5 0 m g /k g ( ip , tw ic e w e e k ly )

M P T P /P +

L A P S -H M T 2 1 1 5 .0 3 n m o l/k g (s c , Q W )

***

*T

H+

are

a in

su

bsta

nti

a n

igra

0

2

4

6

8

-s

yn

uc

lein

(n

g/m

l) V e h ic le



M P T P /P +


*****

α-s

yn

uc

lein

(ng

/ml)

0

3 0

6 0

9 0

1 2 0

Po

le t

es

t (T

-to

tal,

s) V e h ic le

M P T P 3 0 m g /k g , Q D

M P T P 3 0 m g /k g , Q D + L ir a g lu t id e 3 0 n m o l/k g , Q D

M P T P 3 0 m g /k g , Q D + H M 1 5 2 1 1 2 .5 n m o l/k g , Q W

M P T P 3 0 m g /k g , Q D + H M 1 5 2 1 1 5 .0 3 n m o l/k g , Q W***

***

***

***

0

3 0

6 0

9 0

1 2 0

Po

le t

es

t (T

-to

tal,

s) V e h ic le

M P T P 3 0 m g /k g , Q D




***

***

***Vehicle

MPTP 30 mg/kg, QD

MPTP 30 mg/kg, QD + HM15211 2.5 nmol/kg, QW


Vehicle

MPTP 25 mg/kg (sc, twice weekly)

+ Probenecid 250 mg/kg (ip, twice weekly)

MPTP/P +

HM15211 5.03 nmol/kg (sc, QW)

0

1

2

3

4

Tra

cti

on

te

st

(sc

ore

0~

3)

V e h ic le



M P T P /P +


*****


*~***p<0.05~0.001 vs. MPTP or MPTP/P by One-way ANOVA

Str

iatu

mS

ub

sta

nti

a

nig

ra

100x

40x 100x

Vehicle MPTP+ HM15211

2.5 nmol/kg

+ HM15211

5.03 nmol/kg

Su

bsta

nti

a

nig

ra

100x

Str

iatu

m

40x

Vehicle MPTP/P+ HM15211

5.03 nmol/kg

100x

Motor function restoring by HM15211

Subchronic PD model

Chronic PD model

0

1

2

3

Tr

ac

tio

n t

es

t (s

co

re

)

***

**

V e h ic le


M P T P 3 0 m p k , ip +


M P T P 3 0 m p k , ip +


***

Tra

cti

on

te

st

(sco

re)

0

1 0

2 0

3 0

4 0

Po

le t

es

t (

T-tu

rn

, s

)

***

***

V e h ic le


M P T P 3 0 m p k , ip +


M P T P 3 0 m p k , ip +


***P

ole

te

st

(T-t

urn

, s

)

0

3 0

6 0

9 0

1 2 0

Po

le t

es

t (T

-to

tal,

s) V e h ic le

M P T P 3 0 m g /k g , Q D



***

***Po

le t

est

(T-t

ota

l, s

)

0

5 0

1 0 0

1 5 0

2 0 0

Ro

taro

d (

fall

ing

la

ten

cy

, s

)

***

***

V e h ic le


M P T P 3 0 m p k , ip +


M P T P 3 0 m p k , ip +


***

Ro

taro

d(f

all

ing

la

ten

cy, s

)

0

3 0

6 0

9 0

1 2 0

Po

le t

es

t (T

-to

tal,

s) V e h ic le

M P T P 3 0 m g /k g , Q D




***

***

***0

3 0

6 0

9 0

1 2 0

Po

le t

es

t (T

-to

tal,

s) V e h ic le

M P T P 3 0 m g /k g , Q D




***

***

***

Vehicle

MPTP 30 mg/kg, QD



0

1

2

3

4

Tra

ctio

n t

es

t (

sc

or

e 0

~3

)

V e h ic le



M P T P /P +


*****

Vehicle



MPTP/P +


0

1

2

3

4

Tra

cti

on

te

st

(sc

ore

0~

3)

V e h ic le



M P T P /P +


*****

0

2 0

4 0

6 0

Po

le t

es

t (T

-to

tal,

s) V e h ic le



M P T P /P +


***

***

0

5 0

1 0 0

1 5 0

2 0 0

Ro

ta

ro

d (

fa

llin

g l

ate

nc

y,

s)

V e h ic le



M P T P /P +


*** ***

Tra

cti

on

te

st

(sco

re 0

~3

)

Po

le t

est

(T-t

ota

l, s

)


Ro

taro

d(f

all

ing

la

ten

cy, s

)


Anti-inflammatory effect of HM15211

Subchronic PD model

Chronic PD model


200x_Cropped

Vehicle MPTP + HM15211

2.5 nmol/kg

Str

iatu

m

200x

+ HM15211

5.03 nmol/kg

Iba1

+ a

rea

in

str

iatu

m

(% v

s.

ve

hic

le)

0

5 0

1 0 0

1 5 0

2 0 0

Iba

+ a

re

a i

n s

tria

tum

(%

vs

. v

eh

icle

)

V e h ic le


M P T P 3 0 m p k , ip +


M P T P 3 0 m p k , ip +


* * ** * *

* *

0

5 0

1 0 0

1 5 0

2 0 0

IFN

- (

pg

/ml)

***

***

V e h ic le

M PTP 30 m pk, ip

M PTP 30 m pk, ip +

L A P S -H M T 2 1 1 2 .5 n m o l/kg , sc

M PTP 30 m pk, ip +

L A P S -H M T 2 1 1 5 .0 3 n m o l/kg , sc

***

IFN

γ(p

g/m

l)

0

2 0 0

4 0 0

6 0 0

8 0 0

IL-1

0 (

pg

/ml)

*** ***

V e h ic le

M PTP 30 m pk, ip

M PTP 30 m pk, ip +

L A P S -H M T 2 1 1 2 .5 n m o l/kg , sc

M PTP 30 m pk, ip +

L A P S -H M T 2 1 1 5 .0 3 n m o l/kg , sc

***

IL-1

0 (

pg

/ml)

Vehicle MPTP/P

Str

iatu

m

200x

+ HM15211

5.03 nmol/kg

400x

0

5 0

1 0 0

1 5 0

2 0 0

IFN

- (

pg

/ml)

V e h ic le



M P T P /P +


*

0

5 0

1 0 0

1 5 0

2 0 0

2 5 0

3 0 0

3 5 0

Iba

1 +

are

a i

n S

tria

tu

m

(% v

s.

ve

hic

le)

V e h ic le



M P T P /P +


**

*

0

1 0 0 0

2 0 0 0

3 0 0 0

IL-1

0 (

pg

/ml)

V e h ic le



M P T P /P +


******

Iba1

+ a

rea

in

str

iatu

m

(% v

s.

ve

hic

le)

IFN

-γ(p

g/m

l)

IL-1

0 (

pg

/ml)


0

3 0

6 0

9 0

1 2 0

Po

le t

es

t (T

-to

tal,

s) V e h ic le

M P T P 3 0 m g /k g , Q D




***

***

***

0

3 0

6 0

9 0

1 2 0

Po

le t

es

t (T

-to

tal,

s) V e h ic le

M P T P 3 0 m g /k g , Q D




***

***

***Vehicle

MPTP 30 mg/kg, QD



Vehicle

0

1

2

3

4

Tra

cti

on

te

st

(sc

ore

0~

3)

V e h ic le



M P T P /P +


*****

MPTP/P +


0

1

2

3

4

Tra

cti

on

te

st

(sc

ore

0~

3)

V e h ic le



M P T P /P +


*****



0

1

2

3

4

Tra

cti

on

te

st

(sc

ore

0~

3)

V e h ic le



M P T P /P +


*****

Efficacy and related MoAs of Alzheimer’s disease in diabetic mice model


Diabetes / Obesity

Increased insulin resistance

Accumulation of AGE : Vasculature

Impaired glucose metabolism (Peripheral & brain)

Hyperactivation of RAGE

Release of proinflammatory factors

Reactive oxygen species

Cytokines

Worsening of diabetes

Increased risk of Alzheimer’s disease

Accumulation of Aβ, AGE : Brain

12wD0

HM15211 (Q2D for 12 weeks)Sacrifice

db/db ♂6 wks old


Alzheimer’ disease in diabetic mouse model

0

2 0

4 0

6 0

8 0

1 0 0

1 2 0

1 4 0

Am

ylo

id b

eta

1-4

2 (

% v

s.

ve

hic

le)

V e h ic le

H M 1 5 2 1 1 1 .0 8 n m o l/k g

d b /m v e h ic le

d b /d b D 0 (5 w )

***

***

***

0 .0

0 .3

0 .6

0 .9

1 .2

1 .5

1 .8

AG

E (

ug

/ml)

V e h ic le

H M 1 5 2 1 1 1 .0 8 n m o l/k g

d b /m v e h ic le

d b /d b D 0 (5 w )

***

***

Aβ

1-4

2(%

vs

. ve

hic

le)

AG

E (

μg

/ml)

*~***p<0.05~0.001 vs. db/db (18w) vehicle by One-way ANOVA

db/db (18w) vehicle

db/m (18w) vehicle

db/db D0 (6w)

db/db (18w) HM15211 1.08 nmol/kg, Q2D

0

2 0

4 0

6 0

8 0

1 0 0

1 2 0

1 4 0

Am

ylo

id b

eta

1-4

2 (

% v

s.

ve

hic

le)

V e h ic le

H M 1 5 2 1 1 1 .0 8 n m o l/k g

d b /m v e h ic le

d b /d b D 0 (5 w )

***

***

***

0

2 0

4 0

6 0

8 0

1 0 0

1 2 0

1 4 0

Am

ylo

id b

eta

1-4

2 (

% v

s.

ve

hic

le)

V e h ic le

H M 1 5 2 1 1 1 .0 8 n m o l/k g

d b /m v e h ic le

d b /d b D 0 (5 w )

***

***

***0

2 0

4 0

6 0

8 0

1 0 0

1 2 0

1 4 0

Am

ylo

id b

eta

1-4

2 (

% v

s.

ve

hic

le)

V e h ic le

H M 1 5 2 1 1 1 .0 8 n m o l/k g

d b /m v e h ic le

d b /d b D 0 (5 w )

***

***

***

0

2 0

4 0

6 0

8 0

1 0 0

1 2 0

1 4 0

Am

ylo

id b

eta

1-4

2 (

% v

s.

ve

hic

le)

V e h ic le

H M 1 5 2 1 1 1 .0 8 n m o l/k g

d b /m v e h ic le

d b /d b D 0 (5 w )

***

***

***

• Experimental scheme

• Inhibition of Aβ1-42 and AGE accumulation by HM15211

AGE : Advanced Glycated Endproduct

RAGE : Receptor for AGE

Reduction of inflammation and oxidative stress by HM15211

db/m_vehicledb/db _D0 (6w) db/db_Vehicledb/db_HM15211

1.08 nmol/kg

Co

rte

xH

ipp

o_C

A1

Hip

po

_D

G

400x

400x

400x


db/db (18w) vehicle

db/m (18w) vehicle

db/db D0 (6w)

db/db (18w) HM15211 1.08 nmol/kg, Q2D

0

2 0

4 0

6 0

8 0

1 0 0

1 2 0

1 4 0

Am

ylo

id b

eta

1-4

2 (

% v

s.

ve

hic

le)

V e h ic le

H M 1 5 2 1 1 1 .0 8 n m o l/k g

d b /m v e h ic le

d b /d b D 0 (5 w )

***

***

***

0

2 0

4 0

6 0

8 0

1 0 0

1 2 0

1 4 0

Am

ylo

id b

eta

1-4

2 (

% v

s.

ve

hic

le)

V e h ic le

H M 1 5 2 1 1 1 .0 8 n m o l/k g

d b /m v e h ic le

d b /d b D 0 (5 w )

***

***

***0

2 0

4 0

6 0

8 0

1 0 0

1 2 0

1 4 0

Am

ylo

id b

eta

1-4

2 (

% v

s.

ve

hic

le)

V e h ic le

H M 1 5 2 1 1 1 .0 8 n m o l/k g

d b /m v e h ic le

d b /d b D 0 (5 w )

***

***

***

0

2 0

4 0

6 0

8 0

1 0 0

1 2 0

1 4 0

Am

ylo

id b

eta

1-4

2 (

% v

s.

ve

hic

le)

V e h ic le

H M 1 5 2 1 1 1 .0 8 n m o l/k g

d b /m v e h ic le

d b /d b D 0 (5 w )

***

***

***


0

3 0

6 0

9 0

1 2 0

IL-1

(p

g/m

l)

V e h ic le

H M 1 5 2 1 1 1 .0 8 n m o l/k g

d b /m v e h ic le

d b /d b D 0 (5 w )

*

**

IL-1

β(p

g/m

l)

0

1 0

2 0

3 0

4 0

5 0

6 0

7 0

IFN

-

(p

g/m

l)

V e h ic le

H M 1 5 2 1 1 1 .0 8 n m o l/k g

d b /m v e h ic le

d b /d b D 0 (5 w )

***

**

IFN

-γ(p

g/m

l)

0

3

6

9

1 2

1 5

HN

E p

ro

te

in a

dd

uc

t (u

g/m

l)

V e h ic le

H M 1 5 2 1 1 1 .0 8 n m o l/k g

d b /m v e h ic le

d b /d b D 0 (5 w )

***

***

***

HN

E p

rote

in a

dd

uc

t (μ

g/m

l)

J Clin Biochem Nutr. 2007 Jul;41(1):18-26

Summary & Conclusion

• In MPTP/Probenecid induced chronic Parkinson’s disease model, HM15211 inhibited the increase of alpha

synuclein, which is the most prominent pathological biomarker of Parkinson’s disease.

• In aged db/db mice, pathological characters of Alzheimer’s disease such as Aβ1-42 and AGE

accumulations were shown. These were reversed by HM15211 treatment.

• These neuroprotective effects of HM15211 are derived from anti inflammatory effect through the altered

cytokine expression and reduced lipid peroxidation.

Based on these results, the novel long-acting GLP-1 / GIP / Glucagon tri-agonist, HM15211 might have

therapeutic potential for neurodegenerative diseases


Please note presentations reporting more information about HM15211:

119-OR : Therapeutic effect of a novel long-acting GLP-1/GIP/Glucagon triple agonist (HM15211) in NASH and fibrosis animal models

500-P: Bone protective effect of a novel long-acting GLP-1/GIP/Glucagon triple agonist (HM15211) in the obese-osteoporosis rodent model

719-P: A novel combination of a long-acting GLP-1/GIP/Glucagon triple agonist (HM15211) and once weekly basal insulin offers improved glucose lowering and weight

loss in a diabetic animal model

Documents

Therapeutic effect of a novel long-acting GLP-1/GIP/Glucagon … · 2018-10-17 · Therapeutic effect of a novel long-acting GLP-1/GIP/Glucagon triple agonist (HM15211) in NASH and