Comparison of Seladelpar and Combinations with Liraglutide or Selonsertib for Improvement of Fibrosis and

NASH in a Diet-Induced and Biopsy-Confirmed Mouse Model of NASH

RESULTS

www.cymabay.com

FRI-329

BACKGROUND AND AIMS

Yun-Jung Choi1, Jiangao Song1, Jeff D. Johnson1, Marc K. Hellerstein2, Charles McWherter1

1CymaBay Therapeutics, Inc., Newark, U.S.A.; 2Nutritional Sciences and Toxicology, University of California, Berkeley, U.S.A.

Pathway Analysis

12-Week Seladelpar treatment in a diet-induced obesity mouse model of NASH:

▪ More pronounced decrease in hepatic fat fraction than liraglutide, selonsertib or

OCA

▪ Led to a decreased NAS

▪ Resolved established bridging fibrosis

▪ Broadest reduction in fibrosis markers such as collagen expression, protein

synthesis and content

▪ Seladelpar is the only agent that reduced liver hydroxyproline content

12-Week Seladelpar + Selonsertib treatment in the mouse NASH model:

▪ Similar effects to seladelpar alone

12-Week Seladelpar + Liraglutide treatment in the mouse NASH model:

▪ Caused a greater decrease in hepatic fat fraction than either agent alone

▪ Principal component analysis of RNA expression suggests differentiated

mechanisms of action between the two agents

This study confirms the anti-NASH and anti-fibrotic effects of seladelpar and also

provides a rationale for combination with seladelpar and a GLP-1 receptor agonist.

Complete resolution of non-alcoholic steatohepatitis (NASH) is challenging due to

the multifactorial etiology of this complex disease. Combining therapies with

complementary mechanisms of action may achieve better outcomes. Seladelpar,

a potent PPAR delta agonist, attenuates multiple pathophysiologic pathways in

NASH mouse models. Here we evaluate seladelpar and its combinations with the

GLP-1-R agonist liraglutide or the ASK1 inhibitor selonsertib in a diet-induced and

biopsy-confirmed mouse model of NASH.

METHODS

Liver Fibrosis

Treatment Groups

CONCLUSION: Seladelpar for NASH

Male C57BL/6J mice were fed a diet high in fat, fructose and cholesterol for 43

weeks (Gubra, Hørsholm, DK). Prior to treatment, mice with histologically

confirmed steatosis (score ≥ 2) and fibrosis (stage ≥ 1) were randomized into

groups and then treated daily for 12 weeks. Seladelpar, liraglutide, selonsertib

and obeticholic acid (OCA, comparator) were tested alone as were combinations

of seladelpar with liraglutide or selonsertib for their effects on fibrosis and NASH

pathology. Deuterated water (D2O) was administered to allow measurement of

hepatic collagen fractional synthesis rate (FSR). Biochemical (ALT, AST, total

triglycerides (TG) and total cholesterol (TC) and hydroxyproline), liver histological

(NAFLD Activity Score (NAS) and fibrosis) and RNAseq analyses were performed.

Week -43 Week -3 Week -1 Day 1 Week 12

+ Steatosis (H&E) ≥ 2

+ Fibrosis (PSR) ≥ 1

+ Collagen 1a1 (IHC)

Amylin Diet (40% Fat: 20% Fructose: 2% Cholesterol) for 55 Weeks

Induce NASH Liver Biopsy Randomization D2O Labeling for 7 Days

Combination treatment used the same dose as single treatment.

Data are presented as Mean SE; P-values: * < 0.05; ** < 0.01; *** < 0.001; **** < 0.0001

Statistical significance (P-value < 0.05) between groups are presented by different letters (a,b,c,d,e)

NASH Vehicle Seladelpar (10 mg/kg, QD, PO)

Liraglutide (0.2 mg/kg, BID, SC)

Selonsertib (30 mg/kg, BID, PO)

OCA (30 mg/kg, QD, PO) CHOW Vehicle

Seladelpar + Liraglutide

Seladelpar + Selonsertib

RESULTS

Food Intake & Body Weight

Lipids & Liver Enzymes

0 10 20 30 40 50 60 70 80 900

5

10

15

20

Food Intake

Treatment (Days)

Fo

od

In

take (

kC

al)

0 10 20 30 40 50 60 70 80 900

10

20

30

40

50

Body Weight

Treatment (Days)

Bo

dy W

eig

ht

(g)

0

20

40

60

80

100

Plasma TG

TG

(m

g/d

L) a

b

d cd cdbc

e

d

0

20

40

60

80

100

120

Liver TG

TG

(m

g/g

liv

er)

a a

d

b b b b

c

0

100

200

300

400

Plasma TC

TC

(m

g/d

L)

a

b b

c c

d

bc

0

2

4

6

8

10

12

14

Liver TC

TC

(m

g/g

liv

er)

a

ab bc bc

cd

e

bcdd

0

100

200

300

400

Plasma ALT

AL

T (

U/L

)

c

a

a

a

b

bcbc

c

0

100

200

300

400

Plasma AST

AS

T (

U/L

)

a a

d

bc

b

a

b

cd

-80

-70

-60

-50

-40

-30

-20

-10

0

10

20

% C

han

ge f

rom

Baselin

e

-57%

-38%

4%

-47%

-15%

a

ab

b

cd

c

-76%

-51%

cd

d

e -25

-20

-15

-10

-5

0

5

Ch

an

ge f

rom

Baselin

e (

%)

-15%-13%

-4%

-10%

a

a

c

b

cd

dcd

e

-20%

-13%

Relative Change in Fat Fraction Absolute Change in Fat Fraction

Liver Fat FractionPre- & Post-Treatment

NAFLD Activity Score (NAS)

0

1

2

3

4

5

6

7

8

Sc

ore

0

1

2

3

4

Sc

ore

0

1

2

3

4

Sc

ore

0

1

2

3

4

Sc

ore

NAS Steatosis

Hepatocellular Ballooning Lobular Inflammation

Liver Steatosis

NASH Vehicle Seladelpar Liraglutide Seladelpar + Liraglutide

Selonsertib Seladelpar + Selonsertib OCA CHOW Vehicle

NASH Vehicle Seladelpar Liraglutide Seladelpar + Liraglutide

Selonsertib Seladelpar + Selonsertib OCA CHOW Vehicle

0

10

20

30

40

Hep

ati

c F

at

Fra

cti

on

(%

)

0

5

10

15

20

25

Fra

cti

on

al A

rea a

t W

eek

12

(%

)

d

a ab

ccc

a a a

bb

c

aba

bab

c

f

a ab

cdde

bc

a aab

bc

d

a a bbcbc

d

ac c

b b

c c

defef

c ccc

-15

-10

-5

0

5

Ch

an

ge

in

Fra

cti

on

al A

rea

(%

)

Morphometry with steatotic area subtraction

a

b ab

bc

cc

a

abbc

ab

cd

a

bc bbc

cdd

a

bb

cc

b

bcdab

dd

a

cdb

bccc

a

bc

cdd

cdd d

c c

c

c

abc abc

bcc

-SMA Col 1a1 Col IV Laminin Sirius Red Galectin-3

0.00

0.05

0.10

0.15

0.20

Liver Hydroxyproline

Hyd

roxyp

rolin

e(m

g/g

liv

er)

c

b

a

ab

c

ab

c

d

0

1

2

3

4

5

New Collagen I Synthesis(FSR, 7 day D2O labeling)

Daily F

racti

on

al S

yn

thesis

Rate

of

Co

llag

en

I (

%)

aaa

bb

c c

c

Pre Post Pre Post Pre Post Pre Post Pre Post Pre Post Pre Post Pre Post0

20

40

60

80

100

Fibrosis Stage

Pro

po

rtio

n o

f M

ice

(%

)

F0

F1

F2

F3

F4

Haemotoxylin & Eosin StainingSirius Red Staining

0

10

20

30

40

COL1A1

Exp

ressio

n L

evel

(RP

KM

)

a

cc c

abb

cc

0

10

20

30

40

COL3A1

a

cdde

e

ab

bc

cde

de

0

2

4

6

8

TIMP1

a

bcb

c

a

a

bcbc

0

2

4

6

8

MMP2

ab

cd

bc

d

a

ab

cdcd

0

50

100

150

200

250

LCAD

Exp

ressio

n L

evel

(RP

KM

)

d

b

d cc

b

d

a

0

50

100

150

200

IDH1

d

bc

aab

dd

bcc

0

20

40

60

80

100

CPT2

cd

a

bc bcd d

a a

0

500

1000

1500

2000

ACOX1

c

a

c ccc

bb

0

10

20

30

40

CD68

Exp

ressio

n L

evel

(RP

KM

)

a

bc

c

d

a

bb

bc

0

2

4

6

8

TGFB1

a

cdd

e

abbc

cdcd

0.0

0.5

1.0

1.5

2.0

CCR2

ab

c

cd

d

a

b

c c

0

1

2

3

4

MCP-1

a

b bc

c

a a

b b

Fibrosis Markers

Steatosis Markers

Inflammation Markers

RNAseq Analysis

Principal Component Analysis

-SMA Staining

NASH Vehicle Seladelpar Liraglutide Seladelpar + Liraglutide

Selonsertib Seladelpar + Selonsertib OCA CHOW Vehicle

Liver Fibrosis by Morphometric Analysis

Liver Fat by Morphometric Analysis

Phase 2b NASH study is ongoing

NCT035515220

1

2

3

4

5

6

7

8NAS

NA

S

Pre Post Pre Post Pre Post Pre Post Pre Post Pre Post Pre Post Pre Post

****

*

****

***

**

*

0

20

40

60

80

100

120NAS Decrease by 2 points

Pro

po

rtio

n o

f M

ice (

%)

0%

82%

36%

17%

8%

0%

ns

**

**********

100%

67%

ns

ns

vs. Seladelpar by ANCOVA (factor: treatment, covariate:pre-biopsy NAS) vs. Seladelpar by Fisher's exact test

Principal component analysis of the 500 most variable genes; (%) indicates explained variance

-40 -30 -20 -10 0 10 20-20

-10

0

10

20

Principal Component 1 (41%)

Pri

nc

ipa

l C

om

po

ne

nt

2 (

24

%)

Chow Vehicle

NASH Vehicle

Seladelpar

Seladelpar + Selonsertib

Seladelpar + Liraglutide

OCA

LiraglutideSelonsertib